February 2022

Summary of Research Center Project*Briefly describe the general plan of your project (Compile in English within 2 pages.)

Center name: Nano Life Science Institute (NanoLSI)（Within 15 words）
Host institution: Kanazawa University
Head of host institution: Koetsu Yamazaki, President of Kanazawa University（Name, Affiliation）
Center director: Takeshi Fukuma, Professor of Kanazawa University（Name, Affiliation）
Administrative director: Yoshihiro Fukumori, Professor of Kanazawa University（Name, Affiliation）
1) Overall Image of Your Center
NanoLSI is well-known for its world-class bio-scanning probe microscopy (SPM) technologies such as
high-speed atomic force microscopy (HS-AFM) developed by Ando, in-liquid frequency-modulation AFM and
3D-AFM developed by Fukuma, and functional scanning ion conductance microscopy (SICM) developed by
Korchev. In addition, we have been developing novel nanoprobe technologies for imaging the structures,
dynamics, and chemical and mechanical property distributions at the surfaces and/or inside living cells.
Attracted by these technologies, many life scientists have visited our institute from different countries toobserve various nanoscale biological phenomena and understand their fundamental mechanisms. Thus,
NanoLSI is well recognized as one of the world’s most influential centers of Bio-SPM collaborations. In
addition to the Bio-SPM researchers, we have internationally top-level researchers in the life sciences,supramolecular chemistry, and computational science. The interdisciplinary collaborations among these four
disciplines are another distinctive feature of our institute. Our life scientists have different expertise in cellbiology (NPCs, exosomes, among other things), pharmaceutical science (drug metabolism), and cancer
development and progression, allowing us to pioneer novel applications in various life science fields.
Meanwhile, we use our knowledge on supramolecular chemistry for producing molecular sensors, integratingthem into a nanoprobe, and visualizing distributions of target molecules around a living cell. Furthermore,
we also use our expertise in computational science for understanding mechanisms of biological phenomena
from the SPM data obtained. Through these activities, we are pioneering new research discipline “nanoprobe
life science.”2) Research Activities
1. Development of novel nanoprobe technologies
Compared with major bio-imaging techniques such as fluorescence microscopy and electron microscopy, BioSPM has distinctive advantages of molecular-scale or sub-10-nm-scale imaging and quantitative nanomechanicalmeasurements. However, the former is possible only for “in vitro” systems constructed on a solid surface and thelatter is impossible for the intracellular components. Thus, the majority of the nanodynamics at the surface and/or
inside of the living cells have remained inaccessible. In this project, we aim to overcome this limitation by
developing novel nanoprobe technologies for visualizing nanoscale structures, dynamics, chemical and mechanicalproperty distributions at the surfaces and/or inside of the living cells. Major projects include the development of(1) 2D/3D nanoendoscopy-AFM for intracellular imaging, (2) HS-SICM/AFM for molecular-scale cell-surface
imaging, (3) SICM for chemical mapping inside and around living cells, (4) molecular sensors and nano-devices
for sensing and manipulation, and (5) modeling, simulation, and AI-based analysis techniques for understanding
the physics behind the Bio-SPM data. Through these efforts, we aim to create a paradigm shift from “in vitro” to“in vivo” Bio-SPM technology. Meanwhile, we also keep updating our world-class Bio-SPM technologies such as
HS-AFM, 3D-AFM, and SICM. For example, we aim to enhance the imaging rate of HS-AFM from 10 fps to 100
fps for visualizing faster protein dynamics. In this way, we will continue to create novel nanoprobe technologies
for visualizing as yet unseen nanoscale biological phenomena.
2. Understanding of basic cellular functions and their cancer-specific abnormalities
So far, Bio-SPM applications have been mostly limited to biophysics research. In this project, we aim to
expand this application range to molecular cell biology and medical science for the development of the“nanoprobe life science” field. To this end, we aim to present impactful applications of bio-SPM techniques
in different life science disciplines. In the past five years, PIs at NanoLSI from different life science fields
explored possible applications of newly developed live-cell imaging techniques and determined to work on
the focused subjects related to (1) intracellular transport through nuclear pore complexes and its correlation
with the infection mechanisms. (2) cell-cell communication via exosomes to induce cancer cell specificresponse and clarify the nanoscale mechanism. (3) cell dynamics altered by anti-cancer drug using theoriginally designed aptamer, (4) dynamic changes in the cell surface structures and mechanics altered by
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Nano Life Science Institute (NanoLSI)cancer progression, (5) visualizing distribution of one of the metabolites around a living cell for understanding
its correlation with cancer-specific metabolism, (6) machine learning classification for single cell-level cancer
diagnosis, and (7) growth factor receptor dynamics at the living cell surfaces for understanding its correlation
with cancer progression.
3. Establishment of “Nanoprobe Life Science” field
We aim to create a world-leading center for international collaborations on nanoprobe life science. So far,
we have established three programs for promoting Bio-SPM collaborations such as the Bio-SPM summer
school, Bio-SPM collaborative research program, and NanoLSI visiting fellows program. Through these
programs, we will perform many collaborative studies with international visitors and clarify nanoscale
mechanisms of various biological phenomena. In this way, we will play a central role in the establishment of
the “nanoprobe life science” field.
3) Interdisciplinary Research
We combine the world-class Bio-SPM and supramolecular chemistry techniques to develop innovative
nanoprobe methods, including nanoendoscopy. With our unique nanoprobe technologies and computational
analysis techniques, we aim to understand fundamental nanoscale mechanisms of basic cell functions and theircancer-specific abnormalities. In this way, we combine our expertise in nanometrology, supramolecular chemistry,computational sciences, and life sciences to establish a novel research field, “nanoprobe life science.” To this end,
we will continue to take various measures such as the NanoLSI T (Transdisciplinary) meeting, Luncheon meeting(webinar), and NanoLSI Colloquium. The proposals stimulated by the top-down and bottom-up measures arefinancially supported by Transdisciplinary Research Promotion Grants.
4) International Research Environment
We will retain five or more foreign PIs and two international satellites and solicit more robust activities by
the foreign PIs at NanoLSI by strengthening collaborative research with domestic PIs and other NanoLSI
researchers. We will place emphasis on the employment of overseas postdoctoral researchers as well as
outreach programs. We will retain the deployment of 60 or more Bio-SPMs and 6 EMs together with an open
facility policy for international visiting researchers. The deployment of experienced and multi-lingual
technicians will also be retained. We will provide multi-layered financial support for young overseas
researchers such as a startup budget, annual basic research budget and transdisciplinary research promotion
grant. URAs’ support will be provided for overseas researchers to obtain KAKENHI and other external researchfunds. Wide-ranging administrative and livelihood support will also be provided for overseas researchers in
English, including detailed explanation of the terms of the employment contract and Japanese tax system,
accompanying during resident registration, accommodation search and interpreting during signing of thecontract with a real estate agent, etc. In addition, support is provided for the families of overseas researchers.
We will hold several international research meetings per year, including the NanoLSI international symposium
and the NanoLSI-iCeMS joint international symposium.
5) Center Management
NanoLSI has been officially positioned in the statutes of Kanazawa University as an independent instituteof the university. This revision of the statutes has endorsed the Center Director’s discretionary power interms of the personnel arrangements, budget execution, and representation of NanoLSI. The Center Director
will take the initiative in developing interdisciplinary research in particular. The Administrative Director will
work in cooperation to realize the Center Director’s vision of NanoLSI management. We will continuously
apply for a research evaluation-dependent annual salary scheme in order to maintain an internationallycompetitive salary. The research-focused scheme for competent researchers, namely the Research
Professorship, will be continuously applied for full-time NanoLSI researchers to exempt them from nonNanoLSI duties and allow them to focus on their own research with maximal effort. We will make further
efforts to hire female scientists at both the PI level and assistant or associate professor levels. For fostering
future generation researchers of NanoLSI, all 27 full-time researchers of NanoLSI who have an educational
assignment will be engaged as educators in the Graduate School of Frontier Science Initiative, Division of
Nano Life Science. In addition, most of the PIs or Jr. PIs will give lectures to other graduate or undergraduate
program students in various departments, helping us to recruit them to the Graduate School. We will provide
a basic scholarship for the graduate students of the Graduate School from Kanazawa University’s own fund.
In addition, the governmental scholarships given to Kanazawa University will place a priority on the graduate
students in the Graduate School. Kanazawa University will conform to its commitment and will proactively
reassess its existing organization and develop system reforms. The promised deployment of 22 tenured or
tenure-track researchers to NanoLSI has been completed and will be sustained.
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February 2022

Research Center ProjectCenter name: Nano Life Science Institute (NanoLSI)（Within 15 words）
Host institution: Kanazawa University
Head of host institution: Koetsu Yamazaki, President of Kanazawa University（Name, Affiliation）
Center director: Takeshi Fukuma, Professor of Kanazawa University（Name, Affiliation）
Administrative director: Yoshihiro Fukumori, Professor of Kanazawa University（Name, Affiliation）（Name, Affiliation）
1) Overall Image of Your Center* Concisely describe your institute’s identity and provide its mission statement as a WPI center.

1. Mission statement
We combine the world-class bio-scanning probe microscopy (SPM) and supramolecular chemistry
techniques to develop “nano-endoscopic techniques” that allow us to directly image, analyze, andmanipulate the nanodynamics of proteins and nucleic acids on the cell surface or inside the cell.
Furthermore, we use these techniques and multi-scale simulation techniques in a complementary fashion
to elucidate various molecular and cellular dynamics by comparing normal and cancer cells. Based on thedeveloped techniques and knowledge gained through this process, we establish a new academic field,“nano-probe life science”, which aims to fundamentally understand and control various life phenomena,
including development, disease and aging.
2. Center identity
The most salient feature of NanoLSI is our world-class bio-SPM techniques for nanoscale live imaging
of molecular and cellular dynamics, which offer distinctive advantages over other bio-imaging techniques.
For example, electron microscopy allows one to image the static ultrastructure of proteins in vacuum, but
not to directly image protein dynamics in solution. Live imaging by fluorescence microscopy visualizes the
positions of fluorescence-labeled molecules, but does not allow direct imaging of dynamic changes in theprotein structure or the positions of unlabeled molecules. To overcome these limitations, Ando developed
the high-speed atomic force microscope (AFM) and made it possible to directly image the dynamic
structural changes of unlabeled protein molecules in solution (PNAS 2001, 785 citations). In addition,
Fukuma developed the liquid-environment frequency modulation AFM (RSI 2005, 313 citations) andachieved true atomic-resolution imaging even in solution. Based on this technique, he also developed
three-dimensional AFM (3D-AFM) (PRL 2010, 302 citations) and made it possible to visualize the 3D
distribution of water and molecular chains at sub-nanometer resolution. Based on these achievements,
Ando, Fukuma and their group members have led progress in the bio-SPM research field. Therefore, weare confident that we have gathered together the world’s best scientists and launched a center of
excellence aiming at creating novel bio-SPM techniques for visualizing nanodynamics of molecules andcells in solution.
To accomplish our mission, NanoLSI has attracted many internationally top-class scientists with awealth of experience in life science, supramolecular chemistry, and mathematical/computational science.
The Cancer Research Institute of Kanazawa University, the only joint usage/research center in Japan solely
focusing on cancer research, has amassed superb achievements in the study of cancer stem cells, tumor
microenvironment, and molecularly targeted therapy, including the identification of critical molecules
involved in leukemia stem cell regulation (Hirao et al., Nature 2010, 454 citations). Kanazawa University
has also garnered global attention for its advances in supramolecular chemistry, such as the developmentof novel pillar[n]arenes (Ogoshi et al., JACS 2008, 1325 citations). Additionally, we have collaborations
with outstanding scientists at prestigious universities, including the Adam Foster’s group at Aalto University,
one of the few research groups in the world with a strong track record in AFM simulation in liquids, andCarsten Beta’s group at the University of Potsdam that has made great strides in the simulation of complex
systems especially on cell motility. The contributions of these scientists will help us to build an in-depth
understanding of life phenomena based on the results of nanoscale experiments.
Our greatest, defining strengths are our many accomplished scientists and research achievements
across all core disciplines necessary for executing our mission—nanometrology, life science,
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supramolecular chemistry, and mathematical/computational science. Another advantage that sets us apartis that our principal investigators (PIs) include relatively young researchers, meaning that we can continue
to lead the new science of “nano-probe life science” also in the future.
2) Research Activities
2) -1 Research field* Write in the target research field(s)* Describe the importance of the target research field(s), including the domestic and international R&D trends in the field(s) and scientificand/or social significance.
* Describe the value of carrying out research in the field(s) as a WPI center (e.g., Japan’s advantages, global impact on science and/or
society, future prospects)* If there are other centers either in Japan or overseas advancing research in fields similar to the center’s field(s), please list them. (upto 5 organizations)

1. Name of the target research field: Nano-probe Life Science
We establish a novel interdisciplinary research field “nano-probe life science”, where we develop andutilize nano-probe techniques for directly imaging, analyzing and manipulating dynamic behaviors of
biomolecules and cells at nanoscale resolution in physiological environments. This new approach will
promote the fundamental understanding of mechanisms underlying diverse life phenomena such as
diseases and aging.
2. Importance of the target research field[Domestic/global trends and key challenges in this field]
To achieve our mission as outlined above, we will seek to address the following challenges in the core
disciplines of nanometrology, supramolecular chemistry, and life science.
(1) Challenges in nanometrology: Nanoscale imaging, analysis, and manipulation of molecular dynamics
at the periphery and interior of cells
Advances in super-resolution fluorescence microscopy have allowed nanoscale imaging dynamics offluorescence-labeled molecules occurring in the intra-/extracellular spaces (Betzig et al., Science, 346(2014) 439); however, this technique does not support direct imaging of the target molecules’ structuralchanges or the position of many unlabeled molecules. Furthermore, fluorescent tags may interfere withbiomolecular functions. The microscopic imaging of living cells with advanced atmospheric scanningelectron microscopy and polymer membranes have begun to emerge (Takaku et al., PNAS, 110 (2013)7633), but this approach offers optimal resolution only in the tens of nanometers, and thus, is insufficient
to reveal detailed molecular dynamics. Moreover, the electron beams can damage the molecules of interest.
The high-speed AFM developed by Ando at NanoLSI has been used to visualize the nanodynamics ofaquaporins on live bacteria having a relatively hard surface (Yamashita et al., JMB 422 (2012) 300), but it
has not yet been successfully used to measure molecular dynamics on the much softer surfaces of
eukaryotic cells. Although scanning ion-conductance microscopy (SICM) has been used to image
endocytosis and to locate ion channels and receptors on eukaryotic cell surfaces (Korchev et al., Science,
327 (2010) 1653), the optimal resolution of this technique is lower than that of AFM and insufficient for
imaging the nanodynamics of receptors and transducers. SICM has been applied in various techniques,
including one that uses a nanopipette to inject substances into specific intracellular nano-regions, as well
as sample collection and analysis (Mirkin et al., PNAS 104 (2007) 11895). However, SICM has not yet beensuccessfully used to inject molecular machines with high controllability for manipulating protein structures
or functions. In the first half of the grant period, we were able to map the distributions of physicalproperties, such as pH and oxygen concentration, in liquids using molecular sensors (Zhang et al. Nat.
Commun. 10 (2019) 5610). However, this technique has not been applied to the mapping of metabolites.
As these advances and limitations illustrate, progress is being made in the visualization of cell-surface
nanodynamics, but a breakthrough technology is needed for direct imaging of intracellular molecular
dynamics. In addition, the development of techniques requiring the fusion of nanometrology withsupramolecular chemistry, whether for intracellular or cell-surface applications, is a challenge that has yet
to be solved.
(2) Challenges in supramolecular chemistry: Controlling the position and orientation of molecularsensors/machines to operate them on nanostructures of interest
Supramolecular chemistry has developed as a field of chemistry for creating new molecular functions
through the design of selective intramolecular interactions. Recent research attention has focused on
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molecular machines, which were the subject of the 2016 Nobel Prize in Chemistry. These nanoscale devices
can rotate, reciprocally move, or perform other actions in response to external stimuli. Examples include
a molecular elevator that moves up and down in response to pH (Stoddart et al., Science 303 (2004) 1845)
and molecular tweezers that open and close in response to light (Aida et al., Nature 440 (2006) 512).
However, these highly functional molecular sensors and machines are yet to be translated into practicalapplications. One barrier is the lack of technology for effectively controlling the position and orientation ofthese functional molecules so that they can operate on nanostructures of interest. Here, SPM techniques
could help by offering abilities such as the conveyance of molecules to specific nanoscale regions using ananopipette, or sub-nanometer-scale positional control of a molecule affixed to a probe tip. The
combination of SPM and supramolecular chemistry promises to pave the way for effectively controlling andoperating functional supramolecules upon intracellular and cell-surface nanostructures of interest.
(3) Challenges in life science: Achieving nanoscale understanding of cell functions and their cancer-specificabnormalities
Recent progress in whole-genome sequencing of cancer cells has led to identification of driver
oncogenes in many types of cancer. Discovery of drugs that selectively inhibit function of driver geneproducts has established the practice and concept of a molecularly targeted therapy of cancer. Despite theprogress of recent medical and pharmaceutical sciences, mechanisms of malignant progression of cancer,
such as drug resistance and metastasis are still insufficiently understood. This is partly because there are
no techniques for real-time imaging of nano-dynamic features of non-labeled cells and molecules involvedin cancer progression. For example, it is almost impossible to understand the process leading up to drug
resistance without directly imaging the changes that take place inside cells and their environment (e.g.,
pH, oxygen concentration, osmotic pressure, and amino acid and sugar distributions). The significance ofinteractions between cancer cells and the microenvironment mediated by growth factors, inflammatory
cytokines, and exosomes has been increasingly recognized, but without direct imaging of the molecularand cellular dynamics, it is extremely difficult to elucidate mechanisms of adhesion, migration, and invasion
peculiar to cancer cells, through understanding the changes occurring inside and on the surface of cancer
cells. Thus, techniques to directly image and analyze molecular cell dynamics at nanoscale resolution area prerequisite to elucidating the unresolved mechanisms of cell functions and their cancer-specificabnormalities.
[Scientific/social significance]
We seek to tackle the aforementioned challenges in nanometrology, supramolecular chemistry, and life
science through research that expands and combines the knowledge and techniques of each of these
disciplines. Instead of just evolving these fields, we will endeavor to pioneer a whole new discipline of“nano-probe life science” that will have enormous implications for the future of science. The knowledge
and techniques emerging from our research will pave the way to a fundamental understanding of various
biological phenomena that will allow their precise control. Such advances hold immense social significanceas they could aid in conquering cancer and other intractable diseases, extending lifespan, and achieving
other important health improvements.
3. Reasons why our research is a good match for the WPI initiative[Japan’s competitive edge]
Japan’s competitive edge in this research area lies in the fact that it hosts the world’s foremost bioSPM techniques and the inspired minds that created them. As noted earlier, bio-SPM is the surest path torealizing nano-endoscopic techniques. The most fundamental and critical performance factors of bio-SPM
are resolution and speed, and two Japanese scientists from our institute, Fukuma and Ando, have led theworld in the ongoing improvement of these factors. Having designed and built their systems from scratch,both scientists have expertise in developing SPM techniques of superior resolution and speed. Having ateam of scientists of such caliber at one institute is an incredibly strong advantage.
[Appeal as global scientific/social challenges](1) Appeal as a scientific challenge
The origins of material properties and various phenomena can be explained in terms of nanoscale
structures formed from atomic and molecular assemblies and their dynamic behaviors. Hence,
understanding and controlling nanoscale structures would empower us to manipulate at will all sorts ofphysical properties and phenomena. This represents the ultimate aim of science—to transcend theboundaries of the established disciplines of physics, chemistry, biology, pharmacy, and medicine—and
forms the core concept of nanotechnology. Ever since the pursuit of nanotechnological research was
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declared a key pillar of the US strategy by President Clinton in 2000, enormous research as well as financial
investments have been made in nanotechnological R&D worldwide. These efforts have blazed new paths
toward humankind’s understanding and control of natural phenomena at the nanoscale. In the early 2000s,
the main focus was on materials and devices, but in the second half of that decade, bioscience became agreater target of exploration. Subsequently, nanoscience encompassed life sciences such as the medical
and pharmaceutical sciences, evolving into one of the greatest attempts in humankind’s unending quest
to expand the borders of science and technology.
(2) Appeal as a social challenge
Initially, we will perform detailed comparisons of nanodynamics between normal and cancer cells using
our innovative nano-probe techniques. This will lead to fundamental understanding of the mechanisms ofbasic cellular functions and their cancer-specific abnormalities. Achieving this goal will enable precise
control of various phenomena involved in cancer, and thus make it possible to overcome this intractabledisease. Clearly, our research target represents a very important social challenge globally as well as
specifically for Japan, where cancer kills roughly one in every three people.
[Future prospects for this field]
We will endeavor to establish the foundation for nano-probe life science by building techniques for andaccruing expertise in the imaging, analysis, and manipulation of life phenomena at the nanoscale. Further
in future, those techniques and expertise promise to deepen our understanding and control not of only
cancer, but also other life phenomena. Our research field is thus anticipated to pave the way for immense
contributions to human health, including lifespan extension and conquering of intractable illnesses suchas cancer, heart disease, neurodegenerative disease, and liver disease.
４．Japanese and overseas institutes focusing on similar fields
1) Center for Biosystems Dynamics Research (BDR), RIKEN, investigates events unfolding in the bodyduring all stages of the life cycle to gain a comprehensive understanding of the biological functions
regulating living system, and apply these findings to develop and advance medicine and diagnostics.
Although their research goals are similar to ours, unlike us they mainly use optical microscopy toachieve them.
2) Institute of Transformative Bio-molecules, Nagoya University (ITbM) develops novel functional
molecules to visualize and control living systems. Although they combine chemistry and biology totackle issues in life sciences as we do, their focus is on animals and plants. In addition, they use
optical microscopy for imaging.
3) Janelia Research Campus, Howard Hughes Medical Institute (HHMI) focuses on basic medicine, withnovel bio-imaging techniques and neuroscience research at the core. In contrast to us, they use
optical microscopy as the main imaging technique.
4) Max Planck Institute for Multidisciplinary Sciences combines basic research in the natural sciences andmedicine. In contrast to us, they use NMR and optical microscopy as core imaging techniques.
5) The Center for Nanophase Materials Sciences (CNMS) in the USA researches nanotechnology in
general and is globally reputed for SPM research. However, their focus is mainly on the materials andenergy fields, not life sciences.
As described above, while there are numerous research institutes aiming to elucidate the principles ofliving systems, their main imaging tool is optical microscopy. While there are many nanotechnology
research centers in the world, none of them is solely focused on the understanding of life phenomena.
We, as the first in the world, established a research institute for understanding living systems using
advanced nano-probe techniques and to create the novel academic field “nano-probe life science”.
2)-2 Research objectives* Describe in a clear and easy-to-understand manner the research objectives that the project seeks to achieve by the end of its grant
period (in 5 years). In describing these objectives, the following points should be articulated in an easily understandable manner: The
kind of research area(s) you plan to cultivate by, for example, fusing various fields. In that process, what world-level scientific and/or
technological issues are you seeking to solve? What will the expected impact of the scientific advances you aim to achieve be onsociety in the future?
* Describe concretely your research plan to achieve these objectives and any past achievements related to your proposal.

1. Exit goals (5 years from now)
[Research goals]
Our institute is committed to achieving the following goals by the conclusion of the five-year period:
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Develop nano-endoscopic techniques enabling direct imaging, analysis, and manipulation of the dynamic
behavior of molecules inside and on cells.
Acquire an accurate understanding of mechanisms of basic cellular functions as well as their cancer-specificabnormalities by using the nano-endoscopic and computer simulation techniques we will develop.
Pioneer the emerging discipline of “nano-probe life science”.
[Global scientific/technological challenges and their social impact]
Over the years, various techniques have been developed with the aim of gaining a fundamental
understanding of the mechanisms underlying a multitude of life phenomena, including diseases and aging,by directly visualizing molecular and cellular dynamics; however, challenges remain. The resolution ofthese challenges will allow a fundamental understanding and manipulation/control of the mechanisms ofthese phenomena. This endeavor has immense significance for society, as it will contribute to humanhealth by conquering intractable diseases and extend life expectancy.
２．Specific plans for research activities and related achievements to date
[Development of technique for measuring nanodynamics on the cell surface and interior](Development of nano-imaging techniques)・Further improvement of high-speed AFM technologies: The
power of high-speed AFM (HS-AFM) has been demonstrated by anincreasing number of imaging studies on biological molecules (Curr.
Opin. Chem. Biol. 51 (2019) 105). As notable examples, structuralmorphologies of membrane proteins (Nature 574 (2019) 132;
Nature 575 (2019) 395), a liquid-like condensate for autophagy
initiation (Nature 578 (2020) 301) and structural dynamics of
intrinsically disordered proteins (Nat. Nanotechnol. 16 (2021) 181)
have been directly visualized. However, the vast majority of
biological processes have not yet been visualized by HS-AFM. In
order to apply HS-AFM technologies to a wider range of biologicalphenomena, further improvement of HS-AFM technologies are thus
necessary. After NanoLSI was established, we improved devices inthe HS-AFM (i.e., cantilever (Fig. 1a), optical beam deflection
system, amplitude detector (Appl. Phys. Lett. 119 (2021) 181602;
Patent application number (PAN): 2021-121704) and Z-scanner(Rev. Sci. Instrum. 93 (2022) 013701; PAN: 2019-120245) toimprove the scanning performance of HS-AFM. Notably, the onlytrace-imaging mode that eliminates the backward scanning in X-axis
was invented (Fig. 1b, Rev. Sci. Instrum. 92 (2021) 033705; PAN:
2020-199938), enhancing the speed performance of HS-AFM ~2.5
times. The combination of developed devices enables us to perform
Fig. 1: (a) Ultra-small cantilever milled
HS-AFM imaging ~10 times faster than before. As we continue toby FIB. (b) Only-trace-imaging mode.
(c) Concave/convex substrate.
improve the scanning performance of HS-AFM, we plan to apply thisfaster HS-AFM to capture highly mobile biomolecules that have so
far been inaccessible (i.e. actin and microtubule binding proteins, membrane proteins, DNA and RNA
binding proteins, and proteins on isolated organelles and living cells). Furthermore, we have beendeveloping the following devices to extend the functions of HS-AFM: (1) An AFM substrate with controlledconcave/convex shapes that mimic the curvature of biological membranes and that induce mechanicalstress on proteins placed on it (Fig. 1c, Front. Immunol. 11 (2020) 520; Issued patent: JP677310), (2)HS-AFM with nano-manipulator enabling us to manipulate a target object during HS-AFM imaging (PAN:
2019-149584), (3) HS-AFM with patch-clamp technique enabling us to simultaneously record theconformational changes and electrophysiological properties of membrane channel proteins (PAN: 2019153689). As we continue to refine these technologies, we plan to apply them to membrane binding proteins,
cytoskeletal proteins and membrane channel proteins, to extract further insight into the functional
mechanism of these molecules.
・Measurement of nanodynamics on cell surfaces: In the cell membrane, various proteins such as
receptors and channel proteins are present that allow intra/extracellular signaling and substance transport
and are often closely related to cancer development and progression. However, conventional techniques
cannot satisfactorily visualize protein dynamics on live-cell surfaces at the nano level. To overcome thislimitation, we are developing AFM and SICM techniques for visualizing molecular scale nanodynamics on
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a living cell surface.
SICM can image the surface physical properties of
specimens, such as topography, surface charge density, andviscoelasticity, in ionic solutions without mechanical probesample contact. This unique capability is advantageous formeasuring fragile cells, but its maximum imaging rate andspatial resolution are far lower than those desired in biologicalstudies. To solve this problem, we have been continuously
improving the spatiotemporal resolution of our SICM systemsuch that it now performs the highest spatiotemporal resolutionof all existing SICM systems (Rev. Sci. Instrum. 2019). We
established a method for measuring the geometry of an SICM
probe tip with a sub-nm resolution (Fig. 2a, Anal. Chem. 2020),
allowing us to perform the qualitative mechanical mapping ofliving cells by SICM. These developments enabled simultaneous
visualization of the dynamic change in topography andFig. 2: (a) High-resolution SICM probe.
(b) Height and stiffness maps of cancer
mechanical properties of living cancer cells (Fig. 2b) andcells with different cancer driver genes.
revealed the variation of mechanical properties of cancer cells(c) Averaged stiffness of measured cells.
depending on cancer driver genes (Fig. 2c, Biomaterials 2022).
In the next five years, we will further improve thespatiotemporal resolution of SICM to extend the capability of SICM from the sub-cellular to inter-cellularlevel to investigate and characterize cell-cell interactions and communications. To this end, we have beendeveloping a method to further sharpen the SICM probe tip and designing an ultra-low-noise current
amplifier with a wide-bandwidth performance. In addition, we plan to develop a machine-learning-based
denoising technique of a transient signal response from the SICM probe to exceed the usual low-noise
limit of the current amplifier. By combining these developments, we will investigate structural dynamics ofplasma membranes related to various cellular processes associated with changes in physical properties,
such as mechanical and electrical properties, at nanometer-scale resolution.
AFM enables observation of intact samples with sub-nm resolution, but there have been no reports ofobservations of living animal cell surfaces with a sub-100 nm resolution. The reason for this is that the cellsurface is very soft and easily fluctuates during AFM observation. To overcome this problem, we havedeveloped a novel method for cell surface AFM imaging using Micro Porous Silicon Nitride Membrane(MPM), a Si3N4 membrane with 100-200 nm thickness and 1-5 µm diameter holes. In this method, we
culture cells on the MPM and image the bottom cell surface through the holes. As the membrane aroundthe hole is supported by the MPM, the membrane fluctuation and molecular diffusion are suppressed. As
a result, protrusions of less than 10 nm in diameter were successfully imaged on the living colon cancer
cell surface. Furthermore, to recognize specific molecules in the AFM images, we superimposed the AFM
images onto the images obtained using the stimulated emission depletion (STED) microscope andconfirmed that the localization of the E-cadherin molecule observed in the STED images corresponded tothe protruding structures on the cell surface observed in the AFM images. In the next five years, we will
improve the spatiotemporal resolution by exploring various force detection schemes and enhancing the
response speed of various AFM components. We also aim to explore methods to recognize specificmolecules by AFM. Possible candidates include labeling with a structure that can be recognized by AFM,
and functionalizing a tip with a molecular sensor. We plan to apply the method so developed to visualize
clustering behavior of MET receptors on a living cell surface for understanding its correlation with cancer
drug resistance.
・Visualization of intracellular nanodynamics (nano-endoscopic observation): The behavior ofproteins and nucleic acids on the surfaces and in the interiors of organelles such as nuclei and mitochondria
plays important roles in signal transduction through vacuoles and in physiological processes such astransport. Direct visualization of these phenomena at the nano level is unachievable by conventional
techniques. To explore this uncharted realm, we are developing a nano-endoscopic imaging techniquebased on the 3D-AFM technique of Fukuma. In hydration-structure measurement by 3D-AFM, the probe isscanned in a 3D space such that the probe penetrates the hydration structure and the vertical force applied
to the probe apex is recorded to visualize 3D water distribution. Similarly, if we can penetrate themembrane at the cell surface with an acceptable perturbation to cellular activities using a very thin andlong probe, we should be able to scan the probe in a 3D space including the cell interior, for visualizing

Kanazawa University-6
Nano Life Science Institute (NanoLSI

the nanodynamics occurring in the scanned region (Fig. 3a).
So far, we established a method to fabricate a long
nanoprobe (Fig. 3b, Sci. Rep. 2021) and succeeded in
imaging the whole cell structure (Fig. 3c), 3D configurationsof the actin fibers (Fig. 3d), and 2D nanodynamics of theinner scaffold of the bottom plasma membrane (Sci. Adv.
2021). Importantly, our fluorometric assay reveals that such
imaging does not produce detectable changes in cell viability.
Unlike previous AFM techniques using ultrasonic waves or
elastic responses, this method allows an AFM probe todirectly access the target intra-cellular components so that
we can exploit the full range of AFM capabilities such as highresolution imaging, nanomechanical mapping, and molecular
recognition. These features should greatly expand the range
of intra-cellular structures and properties observable in a
Fig. 3: (a) Principle of Nanoendoscopy.
living cell.
(b) Long nanoprobe. (c) Living HeLa
In the next five years, we will continue to optimize thecell. (d) Actin fibers in the live
design and fabrication method of long nanoprobes,
fibroblast cell.
combining nanoendoscopy with various optical microscopy
techniques, and developing analysis techniques for the 2D/3D intra-cellular images. We also aim to
improve the imaging speed and spatial resolution to visualize molecular-scale dynamics, and to expand its
capability from imaging to mechanical property measurements. Meanwhile, we aim to apply this method
for investigating mechanisms of various intra-cellular phenomena, including nucleus stiffness changes
induced by cancer progression or virus infection, formation and disassembling of a focal adhesion, anddifferent cell compaction behaviors induced by E-cadherin variation.
(Development of nano-endoscopic analysis and manipulation techniques)
Based on the nano-endoscopic imaging technique, nano-endoscopic analysis and manipulation techniques
will be developed.
・Injection and sampling of substances using a nanopipette: If substances can be injected into
specific near-surface or intracellular nano-regions, nanodynamics induced by the injection can bemeasured directly using nano-imaging techniques. To date, there are only a limited number of researchgroups who have successfully injected substances into intra/extracellular nano-regions using nanopipettes
and examined the reactions by fluorescence microscopy. The Korchev group is one of these groups. We
combine this technique with the nano-endoscopic imaging technique to enable direct visualization of
nanodynamics that cannot be imaged by fluorescence microscopy. Moreover, sampling from specific nanoregions, which will allow high-precision analysis of the molecules and ions present, has recently become
possible. The Takahashi group has already achieved local sampling of cell cytoplasm for mRNA analysis.
The combination of these leading nanopipette and nano-imaging techniques will make nano-endoscopic
analysis possible.
We established a chemical injection system using nanopipettes over the last five years. The previous
applications of this nanopipette-based chemical delivery or stimulation were limited to the cell surface. In
the next 5 years, we will focus on developing organelle (mitochondria, ER, Golgi body) level local
stimulation technology to examine organelle-organelle interaction. We also investigate cell migration bycontinuous release of chemical substances to cells over several hours using AI-based cell recognition andnanopipette position control. Using the local sampling nanopipette technology, we also focus on organelle
collection to analyze mRNA within the organelle. We have already established a single organelle collection
method with a SICM-fluorescence microscope hybrid system, and we will continue to develop thetechnology for analyzing the collected organelles. Tools to quantify the sample will also be exploited
through combination with a chemical biology approach.
・ Analysis of nano-distribution of physical properties using a molecular sensor:
Intra/extracellular pH and oxygen concentration are closely related not only to cancer development but
also the function of drugs and drug delivery systems. Conventional techniques including fluorescenceimaging are not necessarily sufficient for analyzing nanoscale distributions of these physical properties.
We aim to solve this problem by combining bio-SPM and supramolecular chemistry techniques. We willdevelop molecular sensors that change their structures in response to pH or oxygen concentration insolution using our expertise in supramolecular chemistry. By attaching the sensor to the end of ananopipette for SICM, distributions can be detected based on changes in the ion current passing throughthe pipette. By combining this nanopipette technique with 3D-AFM, the probe apex can be precisely
Kanazawa University-7
Nano Life Science Institute (NanoLSI

positioned to a specific nano-region.
Until now, we have integrated a range of novel nanoprobebiosensors for oxygen gradients, ROS species, ATP, and pH
with our advanced SICM and applied them to map intracellularand extracellular chemical species with higher spatial andtemporal resolution. For example, we have constructed labelfree zwitterionic-like cross-linked glucose oxidase and poly-Llysine pH-sensitive nanoprobes at the tip of dual-barrel
nanopipettes (Fig. 4a-b). These SICM feedback-controlled pHnanoprobes allowed SICM topography and 3D extracellular pH
mapping of living breast cancer MCF7 cells with pH sensitivity
better than 0.01 units, fast response times down to ~2 ms,
and higher spatial resolution of ~50 nm (Fig. 4c). In anotherexample, carbon-filled nanoprobes with a radius as small as 2
nm were functionalized with Pt as ROS-nanoprobe biosensors,which were applied to detect intracellular ROS of melanoma
Fig. 4: (a)Schematic demonstrating
and melanocytes with high sensitivity (Fig. 4d-g) (Nat.
simultaneously
SICM
imaging
andextracellular pH sensing. (b) Nanoprobe
Commun. 2019). We have also developed a new biosensor forshows pH sensitivity in pH-barrel but not inquantitative detection of 1-methylnicotinamide (1-MNA),
SICM-barrel. (c) SICM & fluorescence
GFP
which is produced by the cancer-associated nicotinamide Nimaging and pHe mapping of CD44
MCF7
methyltransferase (NNMT), even in crude biological samples
cells. (d) Optical image of ROS-nanoprobe(Commun. Chem. 2020). The sensitivity of the biosensor was
intracellular in place in a cell. (e)Voltammograms of penetrating ROSimproved through polymer conjugation. In addition, we have
nanoprobe in melanoma. (f) Current traces
started to develop new sensor molecules that can bind toof ROS-nanoprobe inside (red arrows) andlactate, oligosaccharides, and ions with different charges
outside (blue arrows) a melanoma. (g) ROS
2+
3+(Fe and Fe ions) or measure the temperature to probe thelevels measured with ROS-nanoprobe insideenvironment of cancer cells using nanoprobe technology.
melanoma and melanocytes.
In the next five years, we will continue to improve thesensitivity and selectivity of the developed sensors by optimizing the chemical structures and we will also
newly design selective sensors for chiral molecules such as L- and D-amino acids, which will be introduced
into the nanoprobes. We also aim to further improve the sensitivity and selectivity by integrating many
sensors into a polymer film with a controlled mesh size formed at the nanopipette apex. Furthermore, wewill develop tailor-made enzyme-modified nanopipettes and nanoelectrodes functionalized with chemicalsensors based on a supramolecular chemistry approach. For example, we will use a layer-by-layer
technique to modify a nanopipette with the charged enzymes. In addition, we will use metal-organic
frameworks (MOF) to form a molecular-level membrane at the tip of the SICM nanopipette to trap enzymes
and improve the sensitivity and temporal resolution.
・Nano-manipulation using molecular machines: While the development of SPM imaging techniques
is progressing, tools to provide a stimulus to the target specimens at a nanoscale locality, the so-called“nano-manipulation method”, are also indispensable and in high demand. For this purpose, we employ asystem using molecular machines that can be manipulated by various external stimuli, such as changes intemperature, electric potential or light irradiation. More specifically, we use two approaches to allow thesemolecular machines act on targeted places inside or outside cells.
The first is to utilize molecular machines that can interact with the target proteins, organelles, and cells
directly and physically. For instance, the supramolecular chemistry group have synthesized light-operated
molecular machines in which their structures are altered by light illumination. These machines provide thephysical perturbation against cellular components having sophisticated structure such as cell membranes,
DNA, and organelles. Coupling with the SPM technology in NanoLSI, the spatiotemporal dynamics of thesecomponents in response to perturbation is captured in real time, enabling studies on unexploredmechanisms in biology. Supramolecular chemistry researchers have already synthesized the phototriggered lipid (azo-tab), which will be examined in cell membranes and further extended to the othertargets using HS-AFM. In addition, we aim to attach a molecular machine to the end of a SPM probe (SICM
or AFM). In this case, it should be emphasized that the orientation of the molecular machine in the targeted
intra/extracellular site can be controlled, allowing it to act on a specific site with nano level precision. For
example, an exosome plays a critical role as a machine for regulating cellular functions. Hanayama’s groupis working on how antigen presentation by exosomes differs from that of other immune cells. By
immobilizing exosomes derived from immune cells or engineered exosomes expressing immune molecules
on nanopipettes to allow them to act locally on cells and investigate their responses, they are trying toKanazawa University-8
Nano Life Science Institute (NanoLSI

understand the mechanisms of immune response via exosomes at the nanoscale. Thus, in addition tousing machines produced from synthetic chemistry, the “natural” molecular machines that exist in living
systems are also employed for this concept.
In a second approach, the molecular machine, especially a capsule type, is used to alter theconcentration gradient of bioactive molecules close to the SPM observation area at the nanoscale. The
tools for creating nanoscale concentration gradients of small molecules that bind to target proteins andcellular components are accelerating SPM imaging studies to allow the real-time dynamics of biologicalevents to be captured at the nanoscale. To achieve this, supramolecular chemists produce various typesof capsules enabling the on-demand release of the internal bioactive molecule by external stimuli. For the
next five years, using molecular machines including those that have already been developed (JACS 130(2008) 5022; Nature Chem. 6 (2014) 429; JACS 139 (2017) 4631), these practical and useful tools will be
provided to SPM users who are addressing several biological questions.
(Understanding measurement principles of newly-developed nano-probe techniques and life phenomena
by means of mathematical/computational sciences)
・Modeling and simulation of Bio-SPM experiments: In general, the accuracy and reliability of a
newly-developed measurement method should be verified before it is used for practical applications. Such
verifications typically involve comparison with results obtained by an existing method and/or measurementof model samples with known properties. However, both of these approaches are often difficult for an
atomic- or molecular-scale measurement technique. As an alternative solution, simulation techniques are
becoming increasingly important owing to rapid advances in mathematical/computational sciences.
However, very few research groups have experience in reproducing SPM measurements at theatomic/molecular level in silico and verifying the principles behind them. At NanoLSI, we are developing
methods for modeling, simulation and AI-based analysis to understand the principles of various Bio-SPM
experiments.
In the last five years, we have expanded our machine learning infrastructure to predict electrostatic
potentials and hydration structures, while adapting multichannel experimental data as input and taking
the first steps in autonomous AI instrument operation. We have also extended our molecular modelingtools to the study of complex two-dimensional materials (Nature 2020). In particular, we explored thesensitivity of characteristic electronic structure signatures to the properties of the tip and how the analysis
could further be applied to organic systems (Adv. Funct. Mater. 2021). To advance the quantitativeinterpretation of resolution-limited AFM experiments, we have developed the BioAFMviewer software
platform, which integrates simulated AFM scanning and optimized fitting of atomic protein structures toexperimental images (PLoS Comput. Biol. 2020). In addition, we have developed a method for simulating
the dynamic-mode AFM imaging of surfaces of biopolymer assemblies such as chromosomes (J. Phys.
Chem. C 2020).
In the next five years, we will perform simulation of cell membrane penetration with various long
nanoprobes for understanding the optimal conditions for non-invasive nanoendoscopy experiments (Hall,
Foster). We will also perform simulation of 3D-AFM imaging of simple 3D model structures made of carbon
nanotubes and chromosomes for understanding the imaging mechanisms (Sumikama, Fukuma, Foster).
Furthermore, we will develop a contact mechanics model to estimate tension applied to an intra-cellularfiber from a force versus distance curve measured by directly indenting it by a nanoprobe (Okuda).
・Understanding life phenomena: Regardless of the measurement technique, we obtain informationon the structure and properties of targets through probe–target interaction. Therefore, unless we fully
understand the nature of the probe and interaction, even a clear atomic-resolution AFM image may notprovide a complete understanding of the structure of the target. Simulation and AI-based analysis
techniques help in bridging this gap between the measured data and the real-space model. At NanoLSI,
we are developing such methods of analysis for understanding nanoscale mechanisms of biologicalphenomena from Bio-SPM data.
In the last five years, we performed mathematical modelling and simulations to understand pioneeringinteractive HS-AFM experiments of ATP-less walking of the myosin V molecular motor and provide an
explanation in terms of conformational dynamics and energetics. As a result, we clarified that the classicalmodel emphasizing the importance of ATP-induced chemo-mechanical motions, has to be reexamined. We
also developed a method of analysis for HS-AFM movies based on statistical mechanics. The novel method
was then applied to HS-AFM movies filming repeating associations and dissociations of scorpion toxin to a
K+ channel (Fig. 5) (Sci. Adv. 2019). Combining the results of the analysis with the discrete state Markov
model, the binding dynamics of the toxin to the channel were found to occur solely via the induced-fit
pathway, and the states of the channel at each moment in the HS-AFM movies were successfully classified
as a high- or low-affinity state to the toxin.
Kanazawa University-9
Nano Life Science Institute (NanoLSI

In the next five years, we will develop a method forclassifying AFM images of different drug-resistant cancer
cells using a machine learning approach for single-celllevel cancer diagnosis (Foster). We will also develop amethod for producing a 3D and 4D model of protein
dynamics from experimentally observed HS-AFM images
using data assimilation technique (Flechsig). This enables
a deep molecular understanding of biological processes
beyond what is accessible from Bio-SPM experiments. In
addition, we will develop a method to produce a model ofFig 5: HS-AFM movie and mechanism deduced
X-shaped human chromosomes from their 3D-AFM image
from analysis of the movie based on statisticalmechanics
for understanding the internal structures of chromosomes(Sumikama). Furthermore, we continue to work on
simulations of protein dynamics observed by HS-AFM for understanding their mechanisms. We will apply
molecular simulations combined with simulation AFM to active membrane transporters (ABC transporters),
2D protein crystals (Annexin V), and components of basal membranes (laminin networks) (Flechsig).
Examples in progress include 1) further zooming in on the dynamics of association between phospholipase
A2 and cell membranes using molecular dynamics simulations (Sumikama), and 2) applying statisticalmechanics to the biomolecular dynamics of CaMKII and Na+ channels (Sumikama).
[Nano-level understanding of the mechanisms of basic cellular functions using expertise incancer research](Nano-level understanding of basic cellular functions using expertise in cancer research and innovative
nano-probe techniques）
At NanoLSI, we will image, analyze and manipulate nanodynamics of unlabeled nucleic acids, metabolites,
proteins and organelles in normal and cancer cells using nanoendoscopic technologies. This will allow us
to understand the mechanisms of basic cellular functions and their cancer-specific abnormalities.
The control of intracellular transport is vital for cell growth and differentiation. Nuclear pore complexes(NPCs) are multi-protein turnstiles that regulate nucleo‐cytoplasmic traffic. Wong’s group found that NPC
protein TPR regulated autophagy induction in a brain
tumor (Autophagy 2021). Recently, we further
visualized the native NPC and succeeded in theobservation of single filaments inside the inner ringof nuclear pore by HS-AFM (Biomaterials 2020) andchromatinization of DNA (J Phys Chem Lett 2021).
Moreover, we approached the question as to howvarious viruses invade cells and hijack the NPC
functions to replicate themselves. To answer this, wehave started to reveal the conformational dynamics
of influenza protein hemagglutinin (HA) (Nano Lett
2020) and SARS -CoV-2 spike proteins (J Extracell
Vesicles 2021) via HS-AFM. We are actively
collaborating with several groups within the NanoLSI,
with the support of various organoids, exosome
models and nano-endoscopy and HS-AFM analysis.
Thus, we will try to elucidate the nanoscale Fig. 6: Comprehensive research on cancer
mechanisms of intracellular transport (from the cell progression using nano-endoscopic techniques.
membrane to the nucleus), which will contribute tofuture development of diagnostic and therapeutic strategies against viral pandemics.
Exosomes are small extracellular vesicles containing lipids, proteins, RNAs, etc. and participate in cellcell communications, which contribute to variety of cellular functions and phenomena. Hanayama’s group
found how cancer cell-derived exosomes promote cancer metastasis, angiogenesis, and immune-evasion,and thereby participate in the formation of a microenvironment that promotes cancer progression(Carcinogenesis 2020; Int J Cancer 2021; Cell Death Dis 2021; Front Oncol 2021). To get more detailed
information on the characteristics of cancer cell-derived exosomes, we collaborated with Fukuma’s groupto perform 3D-AFM force mapping, which unexpectedly revealed a malignancy-dependent increase in
exosome stiffness due to changes in the amount of elastic fiber-related proteins (Nanoscale 2021). Thisfinding has led to the proposal of a new hypothesis that malignant cancer cells can become more flexible
Kanazawa University-10
Nano Life Science Institute (NanoLSI

and easier to metastasize through the release of these proteins by exosomes. In the next five years, wewill analyze exosomes before their secretion from the cancer cells and after uptake in the
microenvironment cells. This will be performed locally at the single particle level using nanopipettes totrack characteristic molecular changes to elucidate how cancer cell-specific exosomal functions are loadedin the donor cells and released in the recipient cells.
Epitranscriptomic regulation modulates RNA processing, affecting protein expression and function.
Aberrant A-to-I RNA editing is relevant with the initiation and progression of cancer. Nakajima's group
found that the inhibition of ADAR catalyzing RNA editing can suppress cancer cell proliferation, and canincrease the effects of anti-cancer drugs (Pharmacol Ther 2018). Nakajima has also discovered new roles
for drug-metabolizing enzymes controlling pharmacokinetics and toxicity of anti-cancer drugs (Annu Rev
Pharmacol Toxicol 2022). Since there are no small molecular compounds to inhibit ADAR, Nakajima's groupis working on the development of aptamer molecules, a new modality that is increasingly attracting
attention, which potently and specifically inhibit ADAR. We recently succeeded in visualizing the molecular
dynamics of the binding of the aptamer to ADAR by HS-AFM. We will evaluate the effects of the aptamer
on cancer cell viability and invasion ability as well as cell morphology by using SICM.
In the cancer research field, recent technical progress with tumor-derived organoid cultures allowed
us to create sophisticated preclinical cancer models that recapitulate biological processes of human cancer
in mice. Oshima’s group has succeeded in establishing gastric and colon cancer models by using mouseand human cancer-derived organoids (Cancer Res 2018; PNAS 2019; Nat Commun 2020). Using thesemodel systems, we discovered a novel mechanism of polyclonal metastasis (Nat Commun 2021), andshowed defined nanoscale physical properties of the organoid cell surface by using scanning ion
conductance microscope (SICM) (Biomaterials 2022). Using these organoid models and SICM analysis, wewill try to identify the nanoscale mechanisms of malignant progression of cancer cells, which will accelerate
future development of unique diagnostic and therapeutic strategies against cancer.
Stem cells are defined as cells that have the ability to perpetuate an undifferentiated status throughself-renewal, and develop into mature cells through differentiation. Hirao’s group aims to elucidate theessential nature of stem cells, and has identified that inhibition of cell cycle transition, removable of reactive
oxygen species, and the activation of nutrient starvation signals are important for regulation of thestemness and self-renewal of hematopoietic and leukemia stem cells (Nat Med 2006; Cell Stem Cell 2007;
Nature 2010). Therefore, precise measurement of the redox state and the distribution of nutrients suchas glucose and amino acids in cells will enable us to elucidate essential stem cell properties. In this WPI
program, we have revealed several critical cell fate determinants for normal and malignant hematopoiesis(Cell Stem Cell 2018; Nat Immunol 2019). Among them, we discovered critical metabolites for drug
resistance of malignant cells. Based on these findings, we promoted transdisciplinary projects for thedevelopment of a visualization system for small chemical compounds with SPM technology (Commun Chem
2020), leading to a deep understanding of the pathophysiological roles of metabolites. We will develop anovel imaging system for metabolites with sensors developed at NanoLSI, for mapping the extra- or intracellular distribution of the metabolite at the nanoscale, leading to a deep understanding of the pathologicalroles of cancer-specific metabolites. In addition, comparative analyses between normal and cancer stem
cells would lead to identification of cancer stem cell-specific metabolic abnormality.
Molecular-targeted cancer therapy has changed the concepts and practice of cancer treatment.
However, even when molecular-targeted therapies yield a favorable response, relapse due to drug
resistance is a major problem. Yano’s group has discovered that activation of receptor tyrosine kinases,
such as AXL and IGF-1R, leads to the emergence of drug tolerant cells that underly resistance (NatCommun 2019; Nat Commun 2020). In addition, epithelial-to-mesenchymal transition is an independent
mechanism of drug resistance (Cancer Res 2019). Moreover, measurement of the surface profile, elastic
modules, and adsorption of living tumor cells by AFM revealed that tumor cells with an epithelial-tomesenchymal transition associated with drug resistance had a smoother surface compared with drug
sensitive tumor cells. We will establish cell classification methods using machine learning, which enables
the diagnosis of drug resistant tumor cells at a single cell level by AFM.
Cell membrane signal reception and cellular responses are fundamental processes of tissueorganization and homeostasis. Growth factors and their transmembrane receptors play definitive roles indevelopment, regeneration, and cancer progression. Matsumoto‘s group discovered high-performancecyclic peptides that bind to HGF (hepatocyte growth factor) (Nat Chem Biol 2019) and its transmembrane
receptor MET (Nat Commun 2015; Sci Rep 2018). We applied these peptides to 1) analysis of molecular
dynamics by HS-AFM (Nat Chem Biol 2019), 2) molecular recognition using a peptide-conjugated tip in
HS-AFM (ACS Appl Mater Interfaces 2021), and 3) creation of designer therapeutic proteins (Nat Commun
2021; iScience 2021). We will verify: 1) dynamic mechanisms of HGF-MET interaction and MET activation
Kanazawa University-11
Nano Life Science Institute (NanoLSI

on lipid bilayer and living cells by high-speed/high-resolution AFM, 2) the creation and application ofdesigner proteins by Lasso-Graft technology for diagnosis and therapeutics, and 3) molecular imaging ofcancer metastatic niche formation.
Thus, by collaborations with experts in cell biology, cancer biology and nanotechnology, we hope tobuild a path to future developments in life science and a new era of cancer biology.
2)-3 Project management* Describe the center’s research organization (including its research, support and administrative components) and your concept forbuilding and staffing the organization.
* Describe your concrete plan for achieving the center’s final staffing goal, including steps and timetables.
* If the center will form linkage with other institutions, domestic and/or foreign, by establishing satellite functions, provide the name(s)of the partner institution(s), and describe their roles, personnel composition and structure, and collaborative framework with thecenter project (e.g., contracts to be concluded, schemes for resource transfer).
* If the center will form linkage with other institutions, domestic and/or foreign, without establishing satellite functions, provide thenames of the partner institutions and describe their roles and linkages within the center project.
* List in Appendix the principal investigators who are expected to join the center.

Fig. 7: Organization of NanoLSI

NanoLSI members
For the second five years of the WPI grant period, the Center will be comprised of around 80 persons forresearch, around 17 persons for administration, and around 30 persons for support. Sixteen or moreprincipal investigators including 5 or more foreign PIs will be appointed. These PIs will build a soundfoundation for NanoLSI in advancing research.
Satellite institutions
Satellite Research Centers will be maintained in Europe (the UK) and North America (Canada) in order toaccelerate international joint research, train human resources through exchanges of young scientists, andimprove international visibility.
Center in Europe: Imperial College London, London, UK: Professor Korchev as PI
Center in North America: University of British Columbia, Vancouver, Canada: Professor MacLachlan as PI
Partner institutions
Contract-based agreements on joint research, information exchange, mutual support for human resources
development, and joint use of facilities and equipment will be sustained with the following institutions:i.
RIKEN Center for Biosystems Dynamics Research (BDR)
ii.
Microbiology Research Center for Sustainability, University of Tsukuba (MiCS)
iii.
Institute for Quantum Life Science, National Institutes for Quantum Science and Technology(iQLS)
Kanazawa University-12
Nano Life Science Institute (NanoLSI

iv.
Nikon Solutions CO., LTD.
Educational functions
The graduate school, “Division of Nano Life Science,” fosters excellent graduate students who will carryout their own research independently in an excellent research environment with the latest laboratory
equipment at NanoLSI. The supervisors for this graduate school are all world-class researchers who belong
to NanoLSI. In addition, most of the PIs will give lectures to undergraduate students in various
departments, helping NanoLSI to recruit them to the graduate school division. Furthermore, NanoLSI will
start a new lecture course by Jr. PIs and other young researchers for undergraduate students.
Approach for benefitting from diversity
NanoLSI will employ another female PI, while increasing research staff by at least two female assistant or
associate professors every year. For the fixed-term assistant professors, our basic policy was to employ
young foreign researchers, yet NanoLSI has decided to extend this to include Japanese female researchers.
At the end of FY2023, the proportion of female researchers will be over 20%, while keeping the proportionof foreign researchers near 40%.
a) Principal investigators (full professors, associate professors or other researchers of comparable
standing)
At beginning of
project

At end of FY 2021

Final goal
(Date: March, 2023)

Researchers from within the hostinstitution

12

11

12

Foreign researchers invited fromabroad4

4

4Researchers invited from otherJapanese institutions

0

1

1

Total principal investigators

16

16

17

At end of FY 2021

Final goal
(Date: March, 2024)b) Total members
At beginning of project
Number of persons
Researchers
Overseas
researchers
Femaleresearchers％

Number of
persons49％

81

Number of persons

％

81

8

16

31

38

31

38

6

12

15

19

17

21

Principal investigators

16

Overseas PIs

5

31

5

31

5

29

Female PIs

1

6

1

6

2

12

Other researchers

33

Overseas
researchers
Femaleresearchers

16

17

65

64

3

9

26

40

26

41

5

15

14

22

15

23

Research support staff

8

41

29

Administrative staff

13

17

18

Kanazawa University-13
Nano Life Science Institute (NanoLSI

Total number of people whoform the "core" of theresearch center

70

139

128

2)-4 Securing research funding
Past record* Give the total amount of research funding (e.g., competitive funding) secured by the principal investigators who will join the center
project. Itemize by fiscal year (FY2017-2021).

FY
Amount

2017
702

2018
644

2019
686

2020
626

2021
596

(Million Yen)

Prospects after establishment of the center* Based on the past record, describe the concrete prospects for securing resources that match or exceed the WPI project grant.
* Calculate the total amount of research funding (e.g. competitive funding) based on the percentage of time that the researchers will
devote to research activities at the center vis-à-vis the total time they spend on research activities. Be sure that the prospects(FY2022-2026) are realistically based on the past record.

From FY2017 to FY2020, the external research funds secured by all NanoLSI researchers consisting ofPIs, NanoLSI full-time researchers other than PIs, and associated researchers who work at NanoLSI andother departments of KU are shown below. In each year, the amount of external funds secured exceeded
the WPI subsidies received.

FY2017FY2018FY2019FY2020
FY2021

Number of
NanoLSI
researchers

Allocation amount fromthe host institution

Total amount of external funds
50
72
74
83
81

¥886,045,607¥866,977,892¥1,044,068,024¥1,047,075,206¥1,174,472,752¥573,713,736¥629,295,011¥701,070,766¥711,349,232¥844,208,468

The funding track record of NanoLSI is so far quite strong and its continuation to secure the researchfunding of around one billion yen can be realistically expected.
3) Interdisciplinary Research* Describe why interdisciplinary research is necessary and important in the target field(s) and what new field(s) can be expected to becreated by way of this project. Describe your concrete strategy for advancing such interdisciplinary research.

1. Necessity and significance of interdisciplinary research
We aim to gain a fundamental
understanding of the mechanisms of basic
cellular functions at the atomic or
submolecularlevelsbycomparing
nanodynamics inside normal and cancer
cells. To this end, we will expand andcombine our world-leading bio-SPM andsupramolecular chemistry techniques totake a dramatic technological leap toward
thecreationofnano-endoscopic
techniques for direct imaging, analysis, andmanipulation of nanodynamics of proteins,
nucleic acids, and other molecules insideand on the surface of cells, as well as nanodistributions
of
pH
and
oxygenconcentration. Naturally, this work will
Fig. 8: Strategy for interdisciplinary research at NanoLSI.
require collaborations with life scientists inthe medical and pharmaceutical sciences. Moreover, partnering with simulation experts will be essential
for accurately understanding atomic and molecular dynamics from experimental results. As this illustrates,
Kanazawa University-14
Nano Life Science Institute (NanoLSI

the path toward our goal will require an all-out effort that draws upon the techniques and knowledge ofbio-SPM, supramolecular chemistry, medical and pharmaceutical sciences, and mathematical andcomputational sciences. This work will lead to the establishment of the new interdisciplinary science fieldof “nano-probe life science”.
2. Strategy for interdisciplinary research
In our research, there are two key challenges that are tackled by combining expertise in multiple
disciplines. The strategies for achieving this goal are outlined below.
[Development of nano-endoscopic techniques through the combination of bio-SPM andsupramolecular chemistry techniques]
We are developing nano-endoscopic analysis for visualizing the nano-distribution of pH, oxygen
concentration, and other properties that significantly influence molecular and cellular dynamics, using SPM
probes fitted with highly environmental-responsive molecular sensors. Simultaneously, we are seeking toachieve nano-endoscopic manipulation to stimulate nanodynamics using probes fitted with functional
molecules, offering a high-level of control of functionality. Furthermore, we are developing a mode of
combined measurement whereby molecular machines are delivered to nano-regions and a nano-probe will
be used for live imaging of the molecular and cellular dynamics induced by stimuli from the molecularmachines. Additionally, we are developing long nanoprobes functionalized with anti-fouling molecules forreproducible and non-invasive nano-endoscopic imaging. In this way, we combine the expertise in the two
disciplines, nanometrology and supramolecular chemistry.
[Understanding of mechanisms of basic cellular functions and their cancer-specificabnormalities using nano-endoscopic and simulation techniques]
The success of nanometrology depends on not only the performance of the instruments but also theoperating skills of their users and sample preparation conditions. In NanoLSI, SPM researchers are working
on the optimization of the operating conditions while medical/pharmaceutical researchers are optimizing
the sample preparation procedures. In the meanwhile, simulations necessary for proper interpretation of
the measured results are performed by the following two researchers having different expertise inmathematical/computational sciences: Foster, specialized in the atomistic simulation of molecular dynamics
and in ab initio calculation (Phys. Rev. Lett. 102 (2009) 126807; Phys. Rev. Lett. 93 (2004) 187202), andBeta, focusing on modeling and simulation of complicated biological dynamics such as cell motility (PNAS
117 (2020) 6330; Sci. Adv. 6 (2020) eaaz6153). The comparative examination of ourexperimental/simulation results and development of a nanoscale understanding of cellular functions andtheir cancer-specific abnormalities will involve pooling the knowledge of SPM, medical/pharmaceuticalscience, and mathematical/computational science researchers and repeated discussions.
[Formation of international alliance for “nanoprobe life science”]
We will establish an international alliance for the new research field “nanoprobe life science” to achieve
the following three goals.
(Complementary use of various bio-imaging techniques)
Although nanoprobe technology is unique and powerful, it is not the only tool for bio-imaging. There
are many other technologies that are developing very rapidly. The complementary use of these different
technologies will be a key to achieve fundamental understandings of various life phenomena. To achieve
this goal, we have formed an international alliance with other life science institutes having world-leading
bio-imaging technologies. Tight collaborations through this alliance network will also allow us to keep
monitoring the progress of other bio-imaging technologies and to adjust the plan for the development of
our nanoprobe technologies. In this way, we will ensure that our technologies always stay at the frontierof bio-imaging.
(Complementary use of various expertise in molecular cell biology)
At NanoLSI, we have gathered various experts in each important cancer research area as well as inbasic medical and pharmaceutical science. However, fundamental understanding of basic cellular functions
requires a broad range of expertise so that we may need to expand the range of our expertise. To thisend, we continue to seek for a possibility to expand the international alliance network. For particularly
important research areas, we have already recruited talented researchers to join us as Junior PIs, and willcontinue to do so when it is necessary. The formation of international alliance should help us to meet andfind the right people to join our research team.
(Widespread use of nanoprobe technologies for life science research)
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The ultimate goal of our institute is to achieve nano-level understandings on various life phenomenausing advanced nanoprobe technologies. Considering the wide variety of life phenomena, this goal cannot
be achieved by our institute alone. Thus, we aim to open our nanoprobe technologies to all life science
researchers in the world. However, newly developed methods and instruments are not necessarily usable
by non-experts and often need improvements in usability and applicability. Thus, we are initially opening
our technologies to the limited members in the international alliance so that we can obtained detailed
reports from them. Based on the feedback, we will improve our technologies to establish them as common
tools for nanoprobe life science research.
4) International Research Environment4)-1 System for advancing international research* Describe your concrete plan for building an international research center including the makeup of its foreign researchers,
establishment of oversea satellites, and provision of researcher exchanges. Please include a timetable for this plan.
* Describe concretely your strategy for staffing foreign researchers (e.g., postdoc positions) through open international solicitations.
Describe the procedures you will use to do so.

Retaining five or more overseas PIs and two overseas satellites
Of the 16 current PIs at NanoLSI, 5 are from overseas. One of the five is a full-time researcher at
NanoLSI, while the other four stay at NanoLSI for one month or more a year while being employedpermanently at an overseas research institution. In order to promote joint research with these four
overseas PIs, one assistant professor in NanoLSI is assigned to each of them. In addition, one research
associate in charge of joint research is assigned to each of the two laboratories designated as overseas
satellites at Imperial College London, UK and the University of British Columbia, Canada with the financial
support of NanoLSI. This framework of five overseas PIs and two satellites will be retained for the second
five years of the WPI grant period.
Measures to solicit more robust activities by the foreign PIs at NanoLSI.
All four foreign PIs employed at an overseas institution will secure research space and personnel in the
NanoLSI building and will be engaged in collaborative research with domestic PIs and other NanoLSI
researchers. Some of the results obtained by joint research will be published and featured on the NanoLSI
WEB site. To continue these activities under the COVID-19 outbreak, NanoLSI will hold regular online intergroup meetings (T-meetings) between an overseas PI and a domestic PI. In this way, NanoLSI will promote
publications of the achievements obtained and continue to explore new collaborative research projects.
Employment of overseas postdoctoral researchers to be given high priority
When recruiting postdoctoral researchers internationally, the host institution, KU, has a policy of hiringthem as specially appointed assistant professors whose three-year term can be renewed. In accordancewith this policy, NanoLSI conducts recruitment of postdoctoral researchers through international
announcements, and almost all overseas postdoctoral researchers are hired as specially appointed
assistant professors. The number of postdoctoral researchers hired at the end of FY2021 is 28, and the
proportion of those from overseas is 24, i.e., 86% of 28. NanoLSI will retain the policy to employ overseas
young researchers with priority. All NanoLSI recruitments will be done through open international
solicitation.
Outreach programs for external researchers and development of joint research
NanoLSI will place emphasis on outreach programs for external and overseas researchers. The purpose
is to introduce and disseminate NanoLSI's scanning probe microscope (Bio-SPM) technology to external
researchers in the life sciences, which will lead to joint research. They include the Bio-SPM Summer Schoolfor young researchers for one week, the Bio-SPM Collaborative Research for mid-career researchers forabout two weeks, and the NanoLSI Visiting Fellows Program for senior researchers and their researchgroups for one month. Participants will be selected on the basis of calls for applications. NanoLSI will retain
these outreach programs to invite external and overseas researchers to NanoLSI with its world-level in
Bio-SPM technologies. Despite the COVID 19 outbreak, some remote joint research will be attempted.
４) -2 Establishment of international research environment* Describe your concrete strategy for establishing an international research environment and administration system, and the support
system to be provided for researchers from overseas.
* Describe your strategy, procedure and timing for periodically holding international research conferences or symposiums (at least oncea year).

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* Describe your measures to ensure that top-caliber researchers from around the world can work comfortably in carrying out theirresearch within a competitive international environment.

Sharing of Bio-SPM equipment
One of the features of NanoLSI in terms of equipment is the deployment of 60 or more Bio-SPMs and6 EMs. These instruments are not only used by overseas researchers belonging to NanoLSI, but also by
many overseas researchers who visit NanoLSI through various programs for shared equipment.
Deployment of advanced and multi-lingual technicians
In order to support the maintenance of these Bio-SPM instruments and to assist overseas researchers
who use them under this open facility system, one technician who specializes in AFM and speaks threelanguages, English, Chinese, and Japanese, and one technician who is fluent in English and specializes in
EM will be continuously deployed exclusively for NanoLSI. In addition, one Ph. D. technician and one
technician with a master’s degree will be continuously employed to maintain NanoLSI equipment and assist
visiting researchers. This deployment of four advanced technicians will be retained.
Evaluation-dependent annual salary system
NanoLSI will continuously adopt an evaluation-dependent annual salary system in order to ensure aninternationally competitive salary level. The evaluation will be conducted by the Center Director on thebasis of various evaluation criteria, i.e. research performance and contribution to various activities such as
promotion of transdisciplinary research, outreach programs and development of Bio-SPM technologies
related to establishing NanoLSI.
Multi-layered financial support for young overseas researchers
NanoLSI will provide multi-layered support for young researchers to initiate and conduct
interdisciplinary research and to obtain external grants. First, when a young researcher is employed at
NanoLSI, they will be provided with a startup budget of 1 million yen. After spending the startup budget,a basic research budget of half a million yen will be provided every year. In addition to this budgetary
support, Transdisciplinary Research Promotion Grants of 0.5 to 2 million yen per year will be provided after
joint review by PIs of young researchers’ interdisciplinary research proposals.
URAs’ support
In order for young researchers to obtain external research funds, a URA is assigned to NanoLSI who
supports the preparation of application documents through close consultation. With support by the URA
through individual consultation and elaboration/confirmation of the application form of overseas
researchers, 14 young overseas researchers have so far got their own KAKENHI as of February 2022. For
planning and operating outreach projects for researchers such as the Bio-SPM Summer School, another
URA is assigned to be responsible for prior contact with participating overseas researchers and support
after visiting NanoLSI. The deployment of two URAs for the above functions will be retained.
Administrative support in English
In order to maximize the research focus of overseas researchers belonging to NanoLSI, theadministrative staff members assigned to the NanoLSI will provide support in English. The support is wideranging, including explanation of the terms of the employment contract, personnel rules and the Japanese
tax system, and accompanying the researchers during resident registration, driver's license acquisition or
renewal, accommodation search and interpreting during signing of the contract with a real estate agent,interpreting at the time of private car purchase, interpreting when renting a parking lot, interpreting when
opening a bank account, applying for a credit card, etc. In addition, support is provided for the families ofoverseas researchers, including accompanying and interpreting during hospital visits, for finding a nurseryschool, and introduction to community activity circles.
International research meetings
International research meetings such as the NanoLSI Symposium held each year have been held 22
times from FY2017 to FY2021, either sponsored by NanoLSI only or co-sponsored by NanoLSI and otherresearch institute(s). Regarding cooperation with the other WPI centers, iCeMS and NanoLSI have agreed
to organize a joint international symposium once a year. NanoLSI will organize its own international
symposium once a year and keep on holding several international research meetings per year even online
despite COVID-19.
5) Center Management5) -1 Operational management* Describe the role of the center director.
* Describe the role of the administrative director.

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* Concretely describe your concept for establishing an administrative organization.
* Concretely describe the center’s decision-making system.
* Concretely describe how authority is allocated between the center director and the host institution.
* Concretely describe how the center will adopt a rigorous system for evaluating research and will introduce a system for merit-based
compensation (e.g. annual salary scheme). Please describe your procedures and timing for operationalizing these systems.

Role of Center Director: Creating a new field of academic research
The aim of the Center is to create a new interdisciplinary research field called Nanoprobe Life Science,
which integrates nanometrology, life science, supramolecular chemistry, and computational science inorder to elucidate the mechanisms underlying a variety of vital phenomena. To achieve this goal, theCenter Director will draw on his own background as a leader in various research fields to develop a vision
for this interdisciplinary research. The Center Director will have authority to determine strategies and allother matters necessary for achieving this vision as well as deciding on the Center’s direction.
Role of the Administrative Director: Achieving the Center Director’s vision
Based on the Center Director’s strong initiative, the Administrative Director will work in cooperation
with other administrative staff to realize the Center Director’s vision of NanoLSI management in terms offacilities and equipment, competitive fund-raising, outreach activities to both researchers and the public,
cooperation with overseas satellites and other research institutes, assistance for Jr. PIs, employment andfostering of young researchers, holding international symposia and other research meetings, etc. The
Administrative Director will also work on coordination between NanoLSI and the university headquarters
including execution of the host institution’s commitment and system reforms.
Concept for establishing an administrative organization
Kanazawa University positions the success of NanoLSI as one of the most important aspects ofuniversity management and prioritizes the placement of competent administrative staff in NanoLSI. At
present, 17 administrative staff including administrative director are assigned to the NanoLSI
administrative office, 13 of whom can perform their duties in English. NanoLSI administrative staff
members actively participate in planning and organization of international symposia, promotion of public
relations, improvement of the research environment, and livelihood support for overseas researchers. They
contribute to strengthening NanoLSI by maximizing the research focus of the center’s researchers.
Decision-making system
At the beginning of FY2020, NanoLSI was officially positioned in the statutes of Kanazawa University
as an independent institute of the university. This revision of the statutes has endorsed the Center
Director’s discretionary power in terms of the personnel arrangements, budget execution, andrepresentation of NanoLSI. Alongside the Center Director’s discretionary power, the Future Planning Board
consisting of the Center Director, the Administrative Director and four PIs has a steering role. This board
will assess the progress of NanoLSI regularly and discuss medium- to long-term issues. The Administrative
Director, the Center Director’s secretary and other administrative staff hold a meeting each week, reporting
the progress of major projects and activity plans. The Center is operated by transmitting the instructionsof the Center Director to each of the following working groups: alliance formation, open facilities,
transdisciplinary research promotion, research outreach, researcher development, and facilities
management. To ensure that all members of NanoLSI are well informed about the Center’s policies, afaculty board consisting of PIs and professors is held every month and minutes in English and meeting
materials are shared with all staff.
Allocation of authority between the Center Director and the host institution
The statutes of Kanazawa University, the host institution, authorize the Center Director to havecomplete authority over decisions related to the Center’s research strategy, as well as the acquisition andallocation of budgetary resources, personnel, space, and other research resources as well as all othermatters of NanoLSI. To this end, Kanazawa University conforms to its commitment and will proactively
reassess existing regulations and develop system reforms. Regarding cooperation and the sharing of roles
between the host institution and NanoLSI, a regular meeting with the President, Vice President in chargeof general affairs, finance and facilities, the Center Director, and the Administrative Director is held every
month. Here, reports on the activities of NanoLSI are presented, intentions are unified on how to deal
with important matters in the operation of the Center, and the sharing of roles between Kanazawa
University and NanoLSI in dealing with these issues is decided.
Research evaluation and internationally competitive salary scheme
NanoLSI will continuously apply a research evaluation dependent annual salary scheme in order toensure an internationally competitive salary level. The Center Director evaluates the annual research
progress reports submitted by all NanoLSI researchers, and the evaluation score is multiplied maximally
threefold by the NanoLSI researcher allowance to determine the annual salary for the following year.
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5) -2 Research environment* Concretely describe how equipment and facilities, including laboratory space, will be provided in a manner appropriate for a top worldlevel research center. Include your procedure and timing.
* Concretely describe how the center will provide an environment in which researchers can work comfortably on their research by being
exempted from duties other than research and related educational activities, and how they will be provided adequate staff support tohandle paperwork and other administrative functions. Include your procedure and timing.
* Concretely describe how the center will arrange for its researchers to participate in the education of graduate students.

NanoLSI Research Building and Bio-SPM equipment
The host institution, Kanazawa University, completed the new NanoLSI Research Building in November
2020. The new building has a total floor area of 6840 m2, consisting of a basement and 4 floors above
ground. Researchers in the fields of nanometrology, life science, supramolecular chemistry, andcomputational science form a research core under one roof. The structural features of the new building
include the installation of a dry area (empty moat) that protects the entire building from external vibrations
propagating on the ground, and the installation of floating floors that prevent vibrations transmitted frominside the building at key points. Due to these features, high-precision anti-vibration measures are now
possible that could not be realized at existing facilities. The host institution will provide financial supportof 10 million yen per year to maintain the facilities.
Features of the facility include 60 scanning probe microscopes (SPM) (48 atomic force microscopes,
AFM, and 12 scanning ion conductance microscopes, SICM) and 6 electron microscopes (one transmission
electron microscope, TEM, and five scanning electron microscopes, SEM). Most of these microscopes arelocated in the basement, where there is less vibration, forming the research floor that is the hallmark of
NanoLSI. The electron microscopes are used for observing samples; SEMs are used for processing AFM
probes, and TEM is used for measuring the diameter of nanopipettes for SICMs. In addition to SPMs andEMs, the research environment has been enhanced by an animal room, a treatment room, and three P2level laboratories. Furthermore, a shared chemistry laboratory has been established with a total of 16 draft
chambers with high-speed variable air volume control (Variable Air Volume) with excellent energy-saving
performance.
Research-focused system for NanoLSI full-time researchers
The host institution has a unique research focus system for competent researchers. A Research
Professorship will be given to full-time NanoLSI researchers to exempt them from non-NanoLSI duties andfocus on their own research with maximal effort.
URAs and administrative staff support for NanoLSI researchers
For planning and operating outreach projects for researchers such as the Bio-SPM Summer School,
URAs are assigned to be responsible for prior contact with participating overseas researchers and support
after visiting NanoLSI. For international symposia and other research meetings, some administrative staff
take on the responsibility for planning and organization. In addition, the NanoLSI administrative staff cover
all administrative paperwork and other administrative jobs in order to maximize the research focus of
NanoLSI researchers including those from overseas. The NanoLSI researchers are released fromorganizational work for outreach programs, international symposia and other research meetings as wellas administrative details.
Graduate school “Division of Nano Life Science”
For fostering future generation researchers of NanoLSI, the Graduate School of Frontier Science
Initiative, Division of Nano Life Science was established in FY2020 as an education unit paired with
NanoLSI; it is now in operation. This graduate school attracts more excellent students from Japan andoverseas than the fixed enrollment capacity of 6 students per year. To promote interdisciplinary researchin the Division of Nano Life Science, all 27 full-time researchers of NanoLSI who have an educational
assignment are engaged as educators in the Division of Nano Life Science. In addition, these graduate
students in the division can join training courses organized by NanoLSI such as the Bio-SPM summer school
and the transdisciplinary research promotion grants. As for financial support, a master’s student in thedivision will be provided with 130,000 yen per month (breakdown: scholarship ¥50,000 + RA salary¥80,000). A doctorate student receives ¥180,000 per month (breakdown: scholarship ¥100,000 + RA
salary ¥80,000). These scholarships are provided from Kanazawa University’s own funds. In addition, thegovernmental scholarships given to Kanazawa University such as the WISE Program and the Fellowship
Program for Fostering Top Scientists in Fused Disciplines place a priority on the graduate students in the
Division.

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5) -3 Establishing the center in sync with organizational restructuring* Concretely describe the host institution’s organizational reform that will be synchronized with the establishment of the center.
* Describe measures that will be taken by the host institution to sustain the center’s operation after the WPI funding ends. Also describe
how the host institution will promote the centers autonomy after WPI funding ends and how it will over the mid-to-long term
restructure its existing organization in ways that give the center a permanent place within its organization.

Kanazawa University's organizational restructuring
The host institution, Kanazawa University, states in its third mid-term objectives/mid-term plans(FY2016-FY2021) that the university further promotes strong research such as nanotechnology using
innovative atomic force microscopy technology, innovative material development using supramoleculartechnology and research on cancer metastasis and drug resistance mechanisms, in a systematic andintensive manner. At the same time, KU states that the research implementation mechanism will bestrengthened with the aim of establishing research centers for interdisciplinary research.
Based on the stated objectives and approaches, the university reorganized its existing research institute,
the Cancer Research Institute, and subsequently established NanoLSI in FY2017, the Nanomaterials
Research Institute in FY2018, the Advanced Manufacturing Technology Institute in FY2019 and Advanced
Mobility Research Institute in FY2021. Thus a group of multi-disciplinary research institutes has been
established which are independent from conventional departments of the university. While the university
aims to provide a research base to each of these five research institutes in their own multi-disciplinary
academic field, NanoLSI is particularly positioned as a pioneering, path-breaking institute.
Measures taken by the host institution to sustain the Center’s operation after the WPI fundingends
It is essential to secure tenured posts for NanoLSI researchers to sustain and develop NanoLSI as aninternationally top-level research institute over a long period of time. In the FY2021 site visit for WPIinterim evaluation, President Yamazaki of Kanazawa University stated the intention to deploy twenty-two
tenured or tenure track researchers to NanoLSI together with their tenured posts. These promised
deployments were completed in FY2021. NanoLSI and the university headquarters have agreed to keepon these tenured posts for researchers belonging to NanoLSI while they are involved in graduate orundergraduate education in relevant research fields. These personnel measures will build a permanentand sound foundation for NanoLSI in advancing research for many years.
Based on the agreement of President Yamazaki and the Center Director, 6 active young researchers, Jr.
PIs have been employed. Jr. PIs are key players for promoting interdisciplinary research and are treatedas independent researchers in the same way as PIs with a start-up budget of 10 million yen and personnel
expenses for one subordinate young researcher. The final performance reviews for 6 Jr. PIs will be carriedout in FY2023 and FY2024. Six tenure positions for Jr. PIs have been secured at the President's discretion.
President Yamazaki has also agreed to sustain the administrative office and staff both in quality andquantity where all NanoLSI operations are carried out in English, such as research support, daily life
support, and various communications from the office. As of the FY2021 interim evaluation, 18
administrative staff are assigned to the NanoLSI Administrative Office, 14 of whom can perform theirduties in English. Around 18 deployments of administrative staff will be kept on for the second five years
of WPI support period and beyond.
Securing of the Center’s autonomy after the WPI funding period and beyond
At the beginning of FY2020, NanoLSI was officially positioned in the statutes of Kanazawa University
as an independent institute of the university. With this revision of the statutes, the personnel rights, budget
execution rights, and representation rights of the Center Director of NanoLSI have been established bothin real and institutional terms.

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Director’s Vision
Continuous advances in science and technology have enabled humans to explore diverse realms. We have traveled
across the Earth’s surface, atmosphere, and neighboring regions of space and amassed knowledge of various
phenomena, gaining insights that paved the way for today’s prosperity. Yet, there still are many worlds that we have
only begun to explore—the deep sea, the Earth’s interior, the vast reaches of outer space, and, at the other end of thespectrum, the microscopic universe. Optical microscopy, electron microscopy, scanning probe microscopy (SPM),
and other tools have allowed us to peer into domains invisible to unaided human eye. Our observations of
microorganisms, cells, molecules, and atoms have revealed various phenomena and elucidated their origins andphysical properties. Today, we are even able to characterize phenomena based on knowledge of configurations andinteractions among the smallest units of matter—atoms and molecules. Nevertheless, many corners of the nano world
remain uncharted; science and technology are currently unable to provide an accurate portrait of nanoscale structuresand dynamics. This limitation represents a significant barrier to the further advancement of science and technology.
In the life sciences, determining the behavior of proteins, nucleic acids, and other cellular components—basic
building blocks of the human body—is a key step toward fundamentally understanding and controlling the mechanismsof complex biological phenomena, such as development, disease and aging. However, we lack a detailed grasp of these
dynamics. For example, fluorescence microscopy allows us to visualize specific molecules tagged with fluorescentlabels, but we are unable to determine the position or structural changes of the many unlabeled molecules. Electron
microscopy can be used to examine the static ultrastructure of cells and molecules in a vacuum, but not to observe
nanoscale behavior in physiological solutions. At present, SPM is the only available technology for the direct imagingof the position and structural changes of protein molecules in solution, and thus is promising for approaching challenges
in the life sciences. However, SPM does not yet enable the direct observation of the intracellular protein behavior.
Accordingly, many nanoscale properties of intra/extracellular space have yet to be mapped out, limiting our ability tofundamentally understand biological phenomena, such as disease and aging.
Our institute (NanoLSI: Nano Life Science Institute) will explore
these uncharted realms with the aim of elucidating the mechanisms of
biological phenomena at the atomic and molecular levels (≈nanoscale). To advance this mission, we will create innovativetechnologies for direct imaging, analysis, and manipulation of
intracellular and cell-surface nanodynamics. First, we will develop atool for “nanoendoscopic imaging” that, similar to endoscopic
imaging of the stomach cavity, will enable direct video imaging of
intracellular nanodynamics. This tool will visualize on the nanoscale
not only the dynamics of proteins, nucleic acids, and other molecules,
but also the distribution of chemical properties, such as the pH andoxygen concentration, using highly environmentally responsive molecular sensors. We will seek to achieve on thecellular level something akin to therapeutic endoscopy, which includes endoscopic sampling/analysis of foreign
matter in the body and drug injection. Our “nanoendoscopic manipulation” technology will make it possible toKanazawa University-1

Nano Life Science Institute (NanoLSI)

sample/analyze specific nanoscale substances in cells and to control the function of receptors and other biomolecules
via molecular machines with high-level control functions.
Bringing such innovative technologies to reality will be a major challenge, but is not unrealistic. SPM is promising
for the development of nanoendoscopic technologies. SPM visualizes, at the nanoscale, surface topography and the
distribution of physical properties by scanning with a fine-tipped probe. In addition, the probe can be used to apply
stimuli and for structural manipulations with nanoscale precision. Kanazawa University’s achievements in this field
make it a world leader in technological development and applied research in bio-SPM operable in solutions. I
developed the first method for in-solution nanoscale imaging of the water density and surface potential distributions,and Ando was the first in the world to create an SPM technology for the direct nanoscale visualization of protein
dynamics. These technologies form the foundation for nanoendoscopic observations of the behaviors of proteins,
nucleic acids, water molecules, and ions. Moreover, one of NanoLSI’s members, Korchev, established the basis forscanning ion-conductance microscopy, which makes it possible to inject or sample substances in specific nanoscaleregions inside or outside of cells using a nanoscale pipette. Combined, our technologies provide a platform forrealizing nanoendoscopic manipulation. By leveraging these world-leading bio-SPM technologies and collaborations
among the outstanding researchers who pioneered them, NanoLSI will strive to develop nanoendoscopic
imaging/manipulation technologies.
Capitalizing on our innovative technologies for nanoscale imaging, analysis, and manipulation, NanoLSI willseek to establish a fundamental understanding of the mechanisms of basic cellular functions and their cancer-specificabnormalities. This focus on cancer has clear significance, as overcoming intractable diseases is a major goal insociety, and has considerable research implications. Cancer involves many different types of molecular dynamics,
including stemness (i.e., the ability of cells to proliferate by self-renewal and to differentiate), intra/extracellularsignaling, and genome dynamics, and the elucidation of these dynamics on the nanoscale will provide a fundamental
basis for understanding a wide range of biological phenomena. Furthermore, cancer research proceeds from a model
defining normal and abnormal cells; this comparison can be used to determine the relationship between cellular-level
abnormalities and abnormalities in the underlying molecular dynamics. Since cancer has a long research history, a
relatively deep understanding of the determinants of the onset of cancer has been established. Nevertheless, cancer
has yet to be conquered and accounts for one in three Japanese deaths. This suggests that current technologies are
insufficient for fully understanding the underlying molecules and processes. Our proposed nanoendoscopic
technology could provide insight into these elusive factors, and in so doing could lead to significant breakthroughsin the fight against cancer.
This goal cannot be achieved solely by nanometrological researchers. Naturally, medical/pharmaceuticalscientists specializing in oncology, supramolecular chemists with expertise in molecular sensor-based nanoanalyses
and nanomanipulation, and mathematical/computational scientists who can perform multiscale simulations areneeded. NanoLSI will strive to achieve the aforementioned goals through collaborations among researchers withworld-class track records in various fields. Kanazawa University, endowed with the Bio-AFM Frontier ResearchCenter and the Cancer Research Institute, is known worldwide as a leading institute of nanobio research and cancer
research. We will further build upon and expand this foundation to pioneer unexplored nanoscale realms and thus
evolve into a one-of-a-kind research institute, without parallel anywhere else in the world.
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Host Institution’s Commitment
Date 21 February 2022
To MEXTName of host institution:
Name and title of head of host institution:

Kanazawa University
The President, Koetsu Yamazaki

I confirm that the measures listed below will be carried out faithfully regarding “(Nano Life Science Institute)”
adopted under the World Premier International Research Center Initiative.
<Provision in host institution’s mid-to-long-term plan>・Describe clearly the host institution’s mid-to-long-term plan and how it will position the center within that plan.

The host institution, Kanazawa University (hereafter, KU), states in its Third Medium-term
objectives/Medium-term plans (FY2016-FY2021) that KU further promotes strong research such as
nanotechnology using innovative atomic force microscopy technology, innovative material developmentusing supramolecular technology and research on cancer metastasis and drug resistance mechanisms, in asystematic and intensive manner through the on-campus COE system. At the same time, KU states thatthe research implementation mechanism will be strengthened with the aim of becoming a research centerat the highest international level for interdisciplinary, i.e., fused research.
Based on the Third Medium-term objectives/Medium-term plans, KU reorganized its existing institute, theCancer Research Institute, and subsequently established the Nano Life Science Institute, NanoLSI, in FY2017, theNanomaterials Research Institute in FY2018, the Advanced Manufacturing Technology Institute in FY2019 and theAdvanced Mobility Research Institute in FY2021. Thus, a group of multi-disciplinary, fused research institutes has
been established. In FY2020, KU contributed 650 million yen from its own funds and, together with the budget
from MEXT, the new NanoLSI Research Building was completed. While KU aims to provide a research base at thehighest international level in each of these five research institutes in their own multi-disciplinary, i.e., fusedacademic field, NanoLSI is particularly positioned as a pioneering, path-breaking institute. It is clearly stated in theMedium-term plans that "a system will be constructed and operated in which the autonomous operation of theInstitute (NanoLSI) will be fully demonstrated."
In the Fourth Medium-term objectives/Medium-term plans (FY2022-FY2027), it is clearly stated that KU aims
at "strategic expansion/strengthening of outstanding fields, promotion of interdisciplinary, i.e. fused research,
expansion/strengthening of domestic and overseas networks, and promotion of the formation of a world-classacademic center", for which KU "selects research fields that enhance international presence and establishes an
educational and research environment (special research funds, salaries, etc.) that can acquire excellent researchersand students from Japan and overseas”, and that "KU will promote the formation of the world-class academic
centers by fostering and strengthening outstanding research fields and developing good practice throughout theuniversity."<Concrete Measures>・Describe the concrete measures that the host institution will take to satisfy the following requirements.

1)

How the host institution will support the center’s operation and research activities inaccordance with the FY 2017 Application Guidelines, section “6. Host institution’scommitment.”
Kanazawa University n-1

Nano Life Science Institute (NanoLSI)Taking over the results of the Third Medium-term objectives and plans, and aiming to reinforce KU as
research base at the highest international level based on the Fourth Medium-term objectives and plans, KU
will continue to provide multi-layered and priority support to NanoLSI through its concrete measures during
the second half of the WPI grant period as described below:
Provision of Budgets
With the following budget and financial support, KU as the host institution will provide NanoLSI with
substantial support beyond the commitment level of the host institution as of 2017.
-

KU will contribute 400 million yen per year as personnel costs for researchers belonging to NanoLSI.

-KU will continue to support research expenses of 60 million yen per year through the on-campus COE

system of KU as in the first half of the grant period.
-

Ten million yen per year will be provided for the maintenance of the NanoLSI Research Building.

-

For students belonging to the Graduate School of Frontier Science Initiative, Division of Nano Life

Science, the educational unit that fosters future generations of NanoLSI researchers, 50,000 yen per monthfor a master's student and 100,000 yen per month for a doctorate student will be provided as a scholarshipfrom KU’s own funds. In addition, scholarships for the WISE Program for Nano-Precision Medicine, Science,
and Technology, which was applied for by KU and funded by MEXT, will give priority to students in the
Graduate School. Similarly, a scholarship aimed at improving the financial condition of doctorate students,
the Fellowship for Fostering Top Scientists in Fused Disciplines, which MEXT will provide to KU, will givepriority to students in the Graduate School.
-

Regarding the amount of external funding for NanoLSI, after the number of affiliated researchers

exceeded 70 and the research staffing was almost completed, 1,044,068,024 yen (allocation amount to
NanoLSI, 701,070,766 yen) was secured in FY2019, and 1,027,050,241 yen (allocation amount,
700,988,457 yen) in FY2020. In addition to 16 PIs, 6 young researchers have been employed as Jr. PIs
who will lead the next generation, and an excellent research team is in place. It is expected that externalfunds of around 1 billion yen can be secured each year also in the near future, out of the amount of whicharound 700 million yen will be expected to be allocated to NanoLSI.
Personnel Affairs
By preferentially applying the personnel measures described below to NanoLSI, KU will maintain the
research focus of NanoLSI researchers and provide internationally competitive salary levels.
-

KU will assign and maintain excellent full-time researchers (22 positions) with tenure, who will leadresearch promotion in NanoLSI for a long period beyond that of the WPI grant.
-

By preferentially applying KU’s Research Professor system to full-time researchers belonging to NanoLSI

and exempting them from non-NanoLSI duties, KU will secure the research focus of NanoLSI full-time
researchers.
-

KU will maintain an internationally competitive salary level by applying a salary structure according to

the performance of individual researchers, i.e., combining rigorous performance evaluation and evaluationdependent salaries with NanoLSI's unique research allowance.
-

Regarding the enrollment capacity of students (currently 6 students per year) of the Graduate Schoolof Frontier Science Initiative, Division of Nano Life Science, enrollment will be increased from 6 to about
10 in both the master's and doctorate programs by consolidating the overall graduate school enrollment
capacity of KU.
Kanazawa University n-2

Nano Life Science Institute (NanoLSI)- The number of young researcher positions currently employed by WPI subsidies will be secured
by using NanoLSI's abundant external funding.
Infrastructure development
By maintaining and developing the research infrastructure, KU will secure a world-class research
environment for NanoLSI.
- NanoLSI is already officially positioned in the statutes of KU as an independent institute of KU. At thesame time, the independence of NanoLSI is confirmed by securing the center director’s discretionary poweron personnel, budget and representation of NanoLSI. NanoLSI is also positioned as a permanentorganization backed by KU's long-term strategy.
- The Facilities Department of KU will give priority to supporting the maintenance of the new NanoLSI
research building. For the maintenance of 60 Bio-SPMs and 6 EMs deployed in the new building, a technician
specializing in AFM and a technician specializing in EM will continue to be placed in NanoLSI. Bothtechnicians are able to assist users of the equipment in English. In addition to these two technicians, one
technician with a Ph.D. degree and another with a master’s degree will be assigned to NanoLSI, making a
total of four.
- As support for young and overseas researchers, a URA who supports acquisition of research funds byoverseas researchers, and a URA who plans and operates outreach projects for researchers, such as the
Bio-SPM Summer School, will continue to be assigned.
- The administrative system will be maintained in which all NanoLSI operations, such as research work,
daily life, administrative procedures, and various communications from the office, are carried out in English.
Instead of fixing the administrative staff assigned to NanoLSI, turnover will be assured by personnel
transfer, positioning the NanoLSI Administrative Office as a place for OJT to carry out duties in an English
environment.
- In order to maximize the research effort of researchers, a specialist will be assigned to take charge ofwebsite maintenance, revision of research content on the website and the planning and organization ofresearch meetings such as international symposia.
2)

How the host institution will establish a basic policy for the mid-to-long term direction of its
organization and operation, one that restructures its existing organization in ways that givethe center a permanent place within its organization. Please provide a concrete schedule forcarrying out this organizational restructuring.

-

As described in <Provision in host institution's mid-to-long-term plan>, KU clarifies the basic policy

aiming at the formation of a world-class research center through interdisciplinary research and theorganizational restructuring accompanying the formation of research centers. In KU’s implementing
research promotion measures based on that policy, NanoLSI will contribute to the progress ofinterdisciplinary research and the formation of research centers in KU through the transfer of establishedgood practice.
-

KU revised its statutes in FY2020, positioning NanoLSI as an independent institute alongside otherfaculties, supporting the permanent status of NanoLSI.
3)

Ways in which the host institution will provide support to sustain the center as a world
premier international research center after the WPI grant period ends.

-

In order to maintain and develop NanoLSI over a long period of time as a world-class research institute,
Kanazawa University n-3

Nano Life Science Institute (NanoLSI)it is important to secure excellent full-time researchers with tenure. KU has already completed thedeployment of 22 full-time researchers with tenure, as announced at the WPI Interim Evaluation Site Visit.
KU will maintain this deployment. These full-time researchers are engaged in research at NanoLSI and arealso involved in graduate education or bachelor's degree education in order to contribute to thedevelopment of next-generation researchers.
4)

How will a system be instituted under which the center’s director is able to make substantive
personnel and budget allocation decisions necessary to implementing the center project—asystem, which in practice, will allow the center director autonomy in making decisions
regarding the center’s operation.

-

In order to support the long-term status of NanoLSI and the leadership of the center director, the KU

statutes were revised and NanoLSI became an independent institute in FY2020. This measure clarifies theautonomy of NanoLSI and the personnel, budget execution and representation rights of the Center Directorof the NanoLSI.
5) Support that will be provided to the center director in coordinating with other departments
within the host institution when recruiting researchers for the center, while giving reasonable
regard to the educational and research activities of those departments.

-

In launching NanoLSI in FY2017, KU negotiated with its departments, and by transferring full-time

researchers belonging to those departments to NanoLSI or by concurrent appointment to NanoLSI andthe original department, outstanding researchers in the fields of nanometrology, supramolecular chemistry,
molecular biology, cancer research, pharmacy etc. were successfully gathered together in NanoLSI. Whileboth researchers who have been transferred to NanoLSI and researchers who are concurrently appointed
to other departments secure and share research efforts, those researchers are also involved in educationof the Graduate School of Frontier Science Initiative, Division of Nano Life Science or the graduate schoolof their original departments, as well as having some involvement in the bachelor's degree education with
its perspective for fostering future generation NanoLSI researchers. KU will continue to maintain this policy
and system.
6)

Flexibility that will be given in applying, revising, or outfitting the host institution’s internal
systems as needed for the center to effectively implement new management methods (e.g.
English-language environment, merit-based pay, top-down decision making, cooperation in
graduate education) unfettered by conventional modes of operation.

-

KU positions NanoLSI as an institute responsible for pioneering and path-breaking roles in advancing

system reforms. While NanoLSI has been preparing the research environment for the formation of a worldclass research center, KU has been establishing and realizing a new research management environment
in NanoLSI, by not being bound by conventional university management methods but by flexibly operating
the existing system and giving the President new discretionary powers. These include top-down decision
making by regular top leaders’ meetings, a research-focused system for selected researchers, anevaluation-dependent annual salary scheme, a Jr. PI program for the young and brightest, administrative
support in English for overseas researchers, graduate education supported by the research institute, etc.
Based on these results, KU will extend the good practice and know-how established by NanoLSI to otheron-campus research institutes and selected research groups.
7) The accommodation that will be given the center’s infrastructural requirements (for facilities,
Kanazawa University n-4

Nano Life Science Institute (NanoLSI)

e.g., laboratory space; equipment; land, etc.).

-

Concerning the research facility, KU completed the new NanoLSI Research Building in September 2020.

The new building has a total floor area of 6840 m2, consisting of a basement and 4 floors above ground.
Researchers in the fields of nanometrology, supramolecular chemistry, computational science, and life
science form a research core under one roof. The structural features of the new building include theinstallation of a dry area (empty moat) that protects the entire building from external vibrations propagatingon the ground, and the installation of floating floors that prevent vibrations transmitted from inside thebuilding at key points. Due to these features, high-precision anti-vibration measures are now possible that
could not be realized at existing facilities.
-

The facility of the NanoLSI building include 60 scanning probe microscopes (SPM) (48 atomic force

microscopes, AFM, and 12 scanning ion conductance microscopes, SICM) and 6 electron microscopes (onetransmission electron microscope, TEM and five scanning electron microscopes, SEM). Most of thesemicroscopes are located in the basement, where there is less vibration, forming the research floor that is
the hallmark of NanoLSI. In addition to SPMs and EMs, the research environment has been enhanced byan animal room, a treatment room and three P2-level laboratories. Furthermore, a shared chemistrylaboratory has been established with a total of 16 draft chambers with high-speed variable air volume
control (Variable Air Volume) with excellent energy-saving performance. In addition, in order to promoteinterdisciplinary research in the above four fields, a shared laboratory without partitions has been
established on every floor. All explanations of how to use the experimental equipment and the reservation
system are online and in English.
-

Regarding the technicians, one expert in AFM who speaks three languages (English, Chinese andJapanese) and one expert in EM who is fluent in English are deployed exclusively for NanoLSI. In addition,
one technician with a Ph.D. degree and another with a master’s degree are involved in the NanoLSI research
environment.
8) Other types of assistance that will be provided to give maximum support to the center inachieving its concepts and objectives and in becoming a world premier international research
center in both name and deed.

-

A monthly leaders' meeting, which is a top-down decision-making body, attended by the President andVice President in charge of general affairs, finance, and facilities (KU side) as well as the Center Director
and the Administrative Director (NanoLSI side) was held during the first half of the WPI grant period. This
will continue during the second half of the WPI grant. These meetings ensure strong and flexible decisionmaking by the Center Director and close cooperation between KU headquarters and NanoLSI.
9)

How the host institution will self-evaluate the good results of the system reforms achieved by
the center and take the initiative in extending them to departments throughout its
organization.

-

KU will establish the Interfaculty Institute for Frontier Sciences and Research Integration (tentativename) in FY2022 by benefitting from the efforts and know-how of NanoLSI to form a research base interms of personnel, budgetary and research infrastructure management system. This organization iscomposed of a group of interdisciplinary research institutes and a collection of selected research groups
with the aim of forming a new world-class research center for interdisciplinary research and contributing
to solving social problems through comprehensive knowledge. While maintaining its status as an

Kanazawa University n-5

Nano Life Science Institute (NanoLSI)independent research institute, NanoLSI will contribute to the propagation of the good practice of NanoLSI
throughout the university in cooperation with this Interfaculty Institute.
- KU is inspired by NanoLSI's efforts for interdisciplinary research, and will start reforming graduate
school education from FY2022. This reform includes (1) mandatory laboratory rotation in the master's
programs, (2) setting of required subjects across graduate schools such as “data science in society 5.0”,
innovation methodology, technology management, etc., (3) gradual implementation of the principle ofadmission to the doctorate programs by qualifying examination in order to convert all graduate schools
into virtually five-year graduate programs, thus aiming to contribute to basic research and to develop
research personnel with doctorate degrees who will contribute to the solution of social problems withcomprehensive knowledge.
10)

If the host institution has already established a WPI center, it should be fully supporting themaintenance and further development of the existing center and be capable, at the same time,of fully supporting the new center.

Not applicable.

Kanazawa University n-6

Nano Life Science Institute (NanoLSI)

List of Principal Investigators
・If the number of principal investigators exceeds 10, add columns as appropriate.
・Place an asterisk(*) by the name of the investigators who are considered to be ranked among the world’s top researchers.
・For investigators who cannot participate in the center project from 1 April 2022, indicate the time that their participation will start in the“Notes” column.
・Include principal investigators affiliated with satellite institutions. Give the name of their satellite institutions in the “Notes” column.

Name

1

2

3

4

5

6

7

8

9

10

Current affiliation(Institution, department,
specialization)

Academic degree

Takeshi Fukuma*

Nano Life Science
Institute, Kanazawa
University

Ph.D.(Doctor ofEngineering)

Noriyuki Kodera*

Nano Life Science
Institute, Kanazawa
University

Ph.D.(Doctor of
Science)

Shigehisa Akine*

Nano Life Science
Institute, Kanazawa
University

Ph.D.(Doctor of
Science)

Katsuhiro Maeda*

Nano Life Science
Institute, Kanazawa
University

Ph.D.(Doctor ofEngineering)

Tomoki Ogoshi*

Graduate School ofEngineering, Kyoto
University / Nano Life
Science Institute,
Kanazawa University

Ph.D.(Doctor ofEngineering)

Nano Life Science
Rikinari Hanayama* Institute, Kanazawa
University
Nano Life Science
Masanobu Oshima* Institute, Kanazawa
University
Nano Life Science
Atsushi Hirao*
Institute, Kanazawa
University
Cancer Research
Kunio Matsumoto* Institute, Kanazawa
University
Seiji Yano*

11

Miki Nakajima*

12

Richard W. Wong*

13

Yuri E. Korchev*

Notes

Ph.D.(Doctor of
Medicine)
Ph.D.(Doctor of
Vaterinary
Medicine)
Ph.D.(Doctor of
Medicine)
Ph.D.(Doctor of
Science)

University Hospital,
Kanazawa University

Ph.D.(Doctor of
Medicine)

Nano Life Science
Institute, Kanazawa
University
Nano Life Science
Institute, Kanazawa
University
Department of Medicine,
Imperial College
London,UK

Ph.D.(Doctor of
Pharmaceutical
Sciences)
Ph.D.(Doctor of
Medicine)
Ph.D.

Imperial College London
Satellite

14

Department ofChemistry, Faculty ofMark J. MacLachlan* Science, The Universityof British
Columbia,Canada

Ph.D.

15

Adam S. Foster*

Department of Applied
Physics, School of
Science, Aalto
University,Finland

Ph.D.

16

Carsten Beta*

Institute of Physics andAstronomy, University of Ph.D.
Potsdam,Germany
Kanazawa University - 1

The University of British
Columbia Satellite

Nano Life Science Institute (NanoLSI)