The 2nd NanoLSI
Symposium in London
Towards Establishment of New Research Field:
Nanoprobe Life Science– Date –Mon. November 19th, 2018

9:00-17:50– Venue –The Cumberland Hotel – London, UK
Sessions and Posters: Tyburn II
Banquet: Ocean Room0

The 2nd NanoLSI Symposium in London–Towards Establishment of New Research Field: Nanoprobe Life Science

PROGRAMME
MC: Kenji Nakashima, Head of NanoLSI Administrative Office, Kanazawa University

OPENING REMARKS
9:00-9:05

9:05-9:10

Yoshihiro Fukumori
Vice President, Kanazawa University,
Deputy Administrative Director, NanoLSI
Takeshi Fukuma
Director, NanoLSISESSION 1 – Chair: Yasufumi Takahashi
9:10-9:40

Nanopipette biosensors for single-cell analysis
Prof Yuri Korchev (Imperial College London, NanoLSI)

9:40-10:00

High-speed atomic force microscopy for nano-scale imaging of biological samples
Assoc. Prof Mikihiro Shibata (NanoLSI)

10:00-10:20

Development of High-Speed Ion Conductance Microscope
Assist. Prof Shinji Watanabe (NanoLSI)

10:20-10:50

Tea Break, Poster Viewing

SESSION 2 – Chair: Rikinari Hanayama
10:50-11:20

Nanoscale SICM/FRET insights into beta-adrenergic receptor distribution and cAMP
compartmentation in cardiac myocytes
Prof Julia Gorelik (Imperial College London)

11:20-11:40

Three-Dimensional Atomic Force Microscopy for Visualizing Distribution of Water andMolecular Chains at Solid-Liquid Interfaces
Prof Takeshi Fukuma (NanoLSI)

11:40-12:00

Probing molecular processes at solid-liquid interfaces
Prof Adam S. Foster (Aalto University, NanoLSI)

12:00-12:10
12:10-13:30

Photo Session
Lunch, Poster Viewing

1

SESSION 3 – Chair: Shigehisa Akine
13:30-14:00

Novel strategies in molecular sensing
Prof Joshua Edel (Imperial College London)

14:00-14:30

Multifunctional Nanoscale Imaging with Nanopipettes
Prof Patrick Unwin (University of Warwick)
Metabolic control of cancer stemness
Prof Atsushi Hirao (NanoLSI)
Cellulose Nanocrystals: Materials for Photonic Structures
Prof Mark MacLachlan (University of British Columbia, NanoLSI)

14:30-14:50
14:50-15:10

15:10-15:40

Tea Break, Poster Viewing

SESSION 4 – Chair: Masanobu Oshima
15:40-16:10

New fluorescence based methods for imaging single molecules

16:10-16:40

Prof David Klenerman (University of Cambridge)
Assembly and membrane perforation by immune proteins

16:40-17:00

Prof Bart Hoogenboom (University College London)
The nuclear pore complex: nanomachine as gatekeeper of molecular transport acrossnuclear membranes
Prof Richard Wong (NanoLSI)

17:00-17:20

Macrocyclic Peptide Technology Targeting HGF and MET receptor
Prof Kunio Matsumoto (NanoLSI)

17:20

Closing Remarks
Prof Yuri Korchev
Imperial College London, NanoLSI

17:50-19:50

BanquetToast:

Seizo Morita (Administrative Director, NanoLSI)

Closing: Takeshi Fukuma (Director, NanoLSI)2

ORAL PRESENTATION

Yuri Korchev (Imperial College London, NanoLSI) ・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・1
Mikihiro Shibata (NanoLSI)・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・3

Shinji Watanabe (NanoLSI)・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・5

Julia Gorelik (Imperial College London)・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・7

Takeshi Fukuma (NanoLSI)・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・10

Adam S. Foster (Aalto University, NanoLSI)・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・12

Joshua Edel (Imperial College London)・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・14

Patrick Unwin (University of Warwick)・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・16

Atsushi Hirao (NanoLSI)・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・19

Mark MacLachlan (Univeristy of British Columbia, NanoLSI) ・・・・・・・・・・・・・・・・・・・・・・・・・・・・21
David Klenerman (University of Cambridge)

・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・23

Bart Hoogenboom (University College London)・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・25

Richard Wong (NanoLSI)・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・27

Kunio Matsumoto (NanoLSI)・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・29

3

4

Yuri Korchev
Affiliation:
Faculty of Medicine, Imperial College London
Nano Life Science Institute (WPI-NanoLSI), Kanazawa University

Contact:
E-mail: y.korchev@imperial.ac.uk

Education:
1983

B.Sc. in Biology & Physiology, University of St Petersburg

1984

M.Sc. in Physiology, University of St Petersburg

1989

Ph.D. in Biophysics and Cytology, Russian Academy of Sciences

Professional Career:
1984 - 1991

Research Scientist, Russian Academy of Sciences, St Petersburg, Russia

1991 - 1992

Wellcome Trust Research Fellow, St George's Hospital Medical School, UK

1992 - 1995

Post-Doctoral Research Fellow, St George's Hospital Medical School, London, UK

1995 - 1997

Post-Doctoral Research Fellow, Charing Cross & Westminster Medical School, UK

1997 - 2001

Lecturer, Faculty of Medicine, Imperial College London, UK

2001 - 2003

Senior Lecturer, Faculty of Medicine, Imperial College London, UK

2003 - 2005

Reader, Faculty of Medicine, Imperial College London, UK

2005 - present

Professor, Faculty of Medicine, Imperial College London, UK

Research Interests: Development and applications of scanning ion conductance microscopy and f nano-FET and nanopore
sensors for biomedical research.

Publications:

1.

Ren, R et al, (2017). Nanopore extended field-effect transistor for selective single-molecule biosensing. Nature comm. 8: 586
https://doi.org/10.1038/s41467-017-00549-w.

2.

U Vivekananda, et al. (2017) Kv1. 1 channelopathy abolishes presynaptic spike width modulation by subthreshold somatic
depolarization. Proc. Natl. Acad. Sci. U.S.A, 114 (9), 2395-2400

3.

Y Zhang, et al. (2016). Spearhead Nanometric Field-Effect Transistor Sensors for Single-Cell Analysis. ACS nano 10, 3214-3221

1

Nanopipette biosensors for single-cell analysis.
Yuri Korchev
Imperial College London, Division of Medicine,London W12 0NN, UK
Nano Life Science Institute (WPI-NanoLSI), Kanazawa University
Molecular Biology has advanced we know much about the individual molecular components that make up living cells
down to the level of the individual atoms. The challenge, however, is to fully understand the functional integration ofthese components. This requires determining how the molecular machines that make up a living cell are organized andinteract together not at the atomic length scale but on a nm scale. To do this we need to develop and applying
nanoscale techniques for the visualization and quantification of cell machinery in real-time and on living cells. This willlead to detailed, quantitative models of sub-cellular structures and molecular complexes under different conditions forboth normal and diseased cells.
This approach ultimately requires the development of novel biophysical methods. We have recently pioneered thedevelopment of an array of new and powerful biophysical tools based on Scanning Ion Conductance Microscopy thatallow quantitative measurements and non-invasive functional imaging of single protein molecules in living cells.
Scanning ion conductance microscopy and a battery of associated innovative methods are unique among current
imaging techniques, not only in spatial resolution of living and functioning cells, but also in the rich combination ofimaging with other functional and dynamical interrogation methods [1]. There are a significant advance to deliver
nanotechnological solutions to biosensing that are affordable, integrated, fast, capable of multiplexed detection andmonitoring, and crucially to offer high selectivity for the specific detection of trace levels of analyte in biological fluids.
Herein, we design a new class of nanometric field-effect-transistor (FET) sensors [2] and dubbed nexFET (nanopore
extended Field Effect Transistor) [3] that combine the advantages of nanopore single molecule sensing, FETs andrecognition chemistry.
The nexFET biosensors, with controllable gate voltage enable higher molecular throughput, enhanced signal-to-noiseand even heightened selectivity via functionalization of the nexFET with an embedded receptor. This is shown for
sensitive and selective detection of an anti-insulin antibody in the presence of its IgG isotype as well as within complex
mixtures such as blood serum. Self-assembled nanoporous sensors at the tip of nanopipette can be used for
simultaneous SICM imaging and chemical imaging and also can be combined with FET sensors.

References:[1]. A Leo-Macias et al. Nanoscale visualization of functional adhesion/excitability nodes at the intercalated disc. (2016).
Nature comm. 7:10342. doi: 10.1038/ncomms10342[2] Y Zhang, et al. Spearhead Nanometric Field-Effect Transistor Sensors for Single-Cell Analysis. (2016). ACS nano 10,3214-3221, DOI: 10.1021/acsnano.5b05211[3] R Ren, et al. Nanopore extended field-effect transistor for selective single-molecule biosensing. (2017) Nature Com.
8, DOI: 10.1038/s41467-017-00549-w

2Mikihiro Shibata
Affiliation:
Nano Life Science Institute (WPI-NanoLSI)
Institute for Frontier Science Initiative, Kanazawa University

Contact:
E-mail: msshibata@staff.kanazawa-u.ac.jp

Education:
2003

B.S. Nagoya Institute of Technology, Department of Materials Science and Engineering, Japan

2005

M.S. Nagoya Institute of Technology, Department of Materials Science and Engineering, Japan2007

Ph.D. from Nagoya Institute of Technology, Japan

Professional Career:
2005-2007

Research Fellow of the Japan Society for the Promotion of Science, DC1, Japan.

2007-2008

Research Fellow of the Japan Society for the Promotion of Science, PD, Japan

2008-2011

Research Fellow of the Japan Society for the Promotion of Science, SPD, Japan

2011-2013

JSPS Postdoctoral Fellow for Research Abroad, Duke University, USA

2013-2015

Postdoctoral Research Fellow, Max Planck Florida Institute for Neuroscience, USA

2015-2016

Postdoctoral Research Fellow, Kanazawa University, Japan

2016-2017

Associate Professor, Kanazawa University, Kanazawa, Japan2017-present

Associate Professor, Nano Life Science Institute, Kanazawa University, Kanazawa, Japan

Scientific Activities:
2005-present

Member of the Biophysical Society of Japan

2014-present

Member of the Biophysical Society

Research Interests: Bio-imaging, High-speed atomic force microscopy, protein dynamics, cell biology
Honors:2009

Early Career Award in Biophysics Society of Japan2018

The Young Scientists’ Prize from The Commendation for Science and Technology by the Minister of
Education, Culture, Sports, Science and Technology

Publications:
1.

M. Shibata*, H. Nishimasu*, N. Kodera, S. Hirano, T. Ando, T. Uchihashi, and O. Nureki*, Nat. Commun., 8: 1430. (2017).

2.

M. Shibata*, T. Uchihashi, T. Ando, and R. Yasuda*, Sci. Rep., 5: 8724. (2015).

3.

M. Shibata, H. Yamashita, T. Uchihashi, H. Kandori, and T. Ando, Nat. Nanotech. 5: 208 - 212 (2010).

3

High-speed atomic force microscopy for nano-scale imaging of biological samples
Mikihiro Shibata
Nano Life Science Institute (WPI-NanoLSI),
Institute for Frontier Science Initiative, Kanazawa University
Structural biology has long been contributing to our understanding of how proteins function by providing their detailed structures at
the atomic level. And yet, the revealed structures have been restricted to static information, limiting the level of our understanding.
This restriction is now removed by high-speed atomic force microscopy (HS-AFM) that allows direct visualization of individual
protein molecules in action at sub-molecular resolution. HS-AFM studies performed in the last few years have provided newmechanistic insight into the functional mechanism of proteins [1]. For example, HS-AFM movies of bacteriorhodopsin (bR), whichfunctions as a light-driven proton pump, clearly showed that, upon illumination, a cytoplasmic portion of bR displaced toward
adjacent molecules at ~ 0.7 nm [2]. Therefore, high-resolution visualizations of protein dynamics are powerful approaches forstudying elaborate biomolecular processes under realistic conditions.
In this study, we applied HS-AFM to visualize the real-space and real-time dynamics of CRISPR-Cas9, which has been widely used fornumerous applications, such as genome editing. The CRISPR-associated endonuclease Cas9 binds to a guide RNA and cleaves
double-stranded DNA with a sequence complementary to the RNA guide. Our HS-AFM movies showed that apo-Cas9 adopts
unexpected flexible conformations. On the other hand, Cas9–RNA forms a stable bilobed structure and interrogates a target site on
the DNA by three-dimensional diffusion. These HS-AFM movies also provided real-time visualization of the Cas9-mediated DNA
cleavage reaction. Notably, the Cas9 HNH nuclease domain fluctuates upon DNA binding, and subsequently adopts an active
conformation, where the HNH active site is docked at the cleavage site in the target DNA. Collectively, our HS-AFM data extend ourunderstanding of the action mechanism of CRISPR-Cas9 [3].
Figure 1 A sequential HS-AFM images of Cas9–RNA–DNA during DNA cleavage reaction.

References:[1] T. Ando, T. Uchihashi, and S. Scheuring, Chem. Rev. 2014, 114, 3120-3188.
[2] M. Shibata, H. Yamashita, T. Uchihashi, H. Kandori, and T. Ando, Nat. Nanotech. 2010, 5, 208-212.
[3] M. Shibata*+, H. Nishimasu*+, N. Kodera, S. Hirano, T. Ando, T. Uchihashi, and O. Nureki*, Nat. Commun. 2017, 8, 1430. (+equal
contribution, *co-corresponding author)

4

Shinji Watanabe
Affiliation:
Nano Life Science Institute (WPI-NanoLSI)
Institute for Frontier Science Initiative, Kanazawa University

Contact:
E-mail: wshinji@se.kanazawa-u.ac.jp

Education:
2006 Ph.D. Graduate School Science and Technology, Niigata University, Japan, Ph. D. (Science)

Professional Career:
2006

JST-CREST Postdoctoral Fellow, Niigata University

2006-2007

JST- SORST Postdoctoral Fellow, Niigata University

2007

Assistant Professor, Department of Physics, Tohoku University

2007-2009

Assistant Professor, Center for the Advancement of Higher Education, Tohoku University

2009-2012

JST-ERATO Postdoctoral Fellow, Tohoku University

2012-2017

Assistant Professor, Bio-AFM Frontier Research Center, Kanazawa University

2017-present

Assistant Professor, Nano Life Science Institute, Kanazawa University

Scientific Activities:
2012-2013

Committee members of JPS-Division 4 (Semiconductor, Mesoscopic System and Quantum Transport)

2018-present

Committee members of JSAP-M&BE (Molecular Electronics and Bioelectronics)

Research Interests: Scanning probe microscopy techniques for visualizing various physical phenomena
Publications:
1. S. Watanabe, and T. Ando, "High-speed XYZ-nanopositioner for scanning ion conductance microscopy" Appl. Phys. Lett. 111,
113106(4 pp), (2017).
2. Satoru Miyamoto. T. Miura, S. Watanabe, K. Nagase, and Y. Hirayama, “Localized NMR Mediated by Electrical-Field-Induced
Domain Wall Oscillation in Quantum-Hall-Ferromagnet Nanowire” Nano Lett. 16(3), 1596 (6 pp), (2016).
3. T. Hatano, W. Kume, S. Watanabe, K. Akiba, K. Nagase, and Y. Hirayama, “Anisotropic nuclear-spin diffusion in double quantum
wells” Phys. Rev. B 91, 115318 (7 pp), (2015).

5

Development of High-Speed Ion Conductance Microscope
Shinji Watanabe
Nano Life Science Institute (WPI-NanoLSI), Kanazawa University, Kanazawa 920-1192, Japan
In biological studies, scanning probe microscopy (SPM) has been utilized to visualize nanometer-scale surface structures of samples
under physiological liquid environments. Atomic force microscopy (AFM) is very popular and powerful SPM due to its excellent
spatio-temporal resolution. Nevertheless, the AFM probe makes contact with the sample surface, and the exerted forces provide
the deformation to very soft samples such as the plasma membranes of live eukaryotic cells, resulting in low-resolution images andserious damage to the samples [1]. Scanning ion conductance microscopy (SICM)[2] is an alternative imaging way based on an
entirely different working principle for visualizing topographic images (Fig. 1). SICM exploits an electrolyte-filled glass pipette(nanopipette) as a probe and an ion current passing through between an electrode inside the nanopipette and another in an
external bath electrolyte. This ion current is very sensitive to the tip-sample surface separation; therefore, SICM can capture
topographic images without any tip-sample contact. However, the temporal resolution of SICM is much lower than that of AFM. In
SICM, thus, it is hard to visualize topographic changes in specimens that occur in under a minute without sacrificing the spatial
resolution. This significantly limits the applicability of SICM in biological studies.
We recently succeeded in improving the temporal resolution of SICM without sacrificing the spatial resolution to capture
morphological changes in biological samples with nanometer-scale resolution [3]. To improve them, we developed several hardware
components of SICM. The tip-scan-type high-speed XYZ-nanopositioner, its undesired vibration damping method, the nanopipette
tip exhibiting a high spatial resolution, and the concentration gradient method of the electrolyte salt solution enhancing the signal
to noise ratio. These developments allow high-speed SICM images which capture morphological changes in cell membranes in aliquid environment with nanometer-scale resolution. We believe that our studies will strongly contribute to extend the SICM
capabilities in biological applications.

References:[1] J. Seifert, J. Rheinlaender, P. Novak, Y. E. Korchev,
and T. E. Schäffer, Langmuir, 31(24), 6807-6813.
[2] P. K. Hansma, B. Drake, O. Marti, S. A. C. Gould, andC. B. Prater, Science 1989, 243, 641-643.
[3] S. Watanabe, and T. Ando, Appl. Phys. Lett. 2017,
111, 113106-1- 113106-4.

Figure 1 Schematics of high-speed SICM equipment.

6

Julia Gorelik
Affiliation:
Imperial College London

Contact:
E-mail: j.gorelik@imperial.ac.uk

Education:
1989

M.Sc., Biology, St Petersburg Teachers' University, Russia

1996

Ph.D., Cell Biology, Russian Academy of Sciences

Professional Career:
1993-1996

PhD student, Institute of Cytology St Petersburg, Russia

1996-1998

Research Scientist, Institute of Cytology St Petersburg, Russia

1998-2005

Post-Doctoral Research Fellow, Imperial College London

2005-2010

Lecturer, NHLI, Imperial College, London

2010-2011

Senior Lecturer, NHLI, Imperial College, London

2011-2014

Reader in Cellular Biophysics, Imperial College London

2014-Present

Professor of Cellular Biophysics, Imperial College London

Research Interests: Interface between physics and cardiovascular research. Scanning Ion Conductance Microscopy (SICM),
functional imaging of the cardiovascular system

Honors:Wellcome Trust University Award
Imperial College Research Excellence Award
Invited Professorship, Pierre and Marie Curie University, Paris,
Invited Professorship, Verona University, Italy,

Publications:
1. Sanchez-Alonso JL, Bhargava A, O'Hara T, Glukhov AV, Schobesberger S, Bhogal NK, Sikkel MB, Mansfield C, Korchev YE, Lyon AR,
Punjabi PP, Nikolaev VO, Trayanova NA, Gorelik J. () Microdomain-Specific Modulation of L-type Calcium Channels Leads to
Triggered Ventricular Arrhythmia in Heart Failure. Circ Res. 2016: 119(8):944-55
2. Glukhov AV, Balycheva M, Sanchez-Alonso JL, Ilkan Z, Alvarez-Laviada A, Bhogal N, Diakonov I, Schobesberger S, Sikkel MB,
Bhargava A, Faggian G, Punjabi PP, Houser SR, Gorelik J. Direct Evidence for Microdomain-Specific Localization and Remodeling ofFunctional L-Type Calcium Channels in Rat and Human Atrial Myocytes. Circulation. 2015: 132(25):2372-84
3. Nikolaev VO, Moshkov A, Lyon AR, Miragoli M, Novak P, Paur H, Lohse MJ, Korchev YE, Harding SE, Gorelik J. {beta}2-Adrenergic
Receptor Redistribution in Heart Failure Changes cAMP Compartmentation. Science. 2010, 327(5973):1653-7

7

Nanoscale SICM/FRET insights into beta-adrenergic receptor distribution and cAMP
compartmentation in cardiac myocytes
Julia Gorelik
Imperial College London
The b1- and b2 adrenergic receptors dominate the cardiac response to catecholamines through their coupling to Gs proteins and to
the production of the common second messenger cAMP (Xiang and Kobilka (2003)). Cyclic AMP controls thecatecholamine-dependent changes in rate, force and speed of contraction of the heart. However, selective stimulation of these two
receptor subtypes leads to clearly distinct physiological and pathophysiological responses. In healthy cardiac myocytes b1- but notb2ARs stimulate the cAMP-dependent protein kinase mediated phosphorylation of phospholamban and cardiac contractile proteins(Xiao (2001)). Heart-specific overexpression of b1- but not of b2AR in transgenic mice induces progressive cardiac hypertrophy andheart failure (Engelhardt et al (1999)). Such subtype specific differences have been attributed to distinct patterns of cAMP
compartmentation from the two receptor subtypes observed using electrophysiological and fluorescent biosensors (Nikolaev et al(2006)). However, the exact distribution of the b1- and b2ARs is still elusive. Unfortunately, methods to detect such localised effects
have so far been limited.
Our new functional approach combines scanning ion conductance microscopy (SICM) with measurements of cAMP production byfluorescence resonance energy transfer (FRET) after localized receptor stimulation (Nikolaev et al (2010)). SICM is a non-opticalmethod which uses a nanopipette as a scanning probe to image the surface topography of living cells and allows to resolve thestructural features of cardiac myocytes such as Z-grooves and transverse (T) tubules with a resolution equal to the pipette’s innerdiameter (Novak et al (2009)), typically ~50 nm. After the acquisition of the cell surface topography we precisely position the pipette
onto various membrane regions of defined morphology to locally apply agonists and antagonists of the bARs. Local stimulation isachieved by applying pressure into the pipette while constantly superfusing the cells with the buffer/antagonist solution from theside (Nikolaev et al (2006)). Upon selective stimulation of b1- and b2ARs we measure cAMP synthesis in defined areas of the cells by
FRET microscopy using a sensitive cAMP sensor Epac2-camps (Nikolaev et al (2004)).
To study the localization of receptors and cAMP signals in cardiomyocytes isolated from healthy adult rat hearts, we locallystimulated b1- and b2ARs either in the T-tubule or on the crest of the cell. Selective stimulation of b1ARs in both regions resulted in arobust decrease of FRET, reflecting the stimulation of cAMP synthesis by the receptors localized in different parts of the membrane.
In sharp contrast, b2AR selective stimulation led to cAMP signals only in the T-tubules, but not in the outer regions of themembrane.
In a rat model of chronic heart failure, β2ARs redistributed from the transverse tubules to the cell crest, leading to diffuse
receptor-mediated cAMP signaling. We showed that surface morphology and T-tubular structure are significantly disrupted inchronic heart failure cells (Lyon et al (2009)), and that impacts on the redistribution of sarcolemmal β2AR and localized secondary
messenger signaling.

References:(1) Engelhardt S, Hein L, Wiesmann F, Lohse MJ. Progressive hypertrophy and heart failure in beta1-adrenergic receptor transgenic
mice. Proc Natl Acad Sci U S A 1999; 96(12):7059-7064.

8

(2) Lyon AR, MacLeod KT, Zhang Y, Garcia E, Kanda GK, Lab MJ Korchev YE, Harding SE, Gorelik J. Loss of T-tubules and other changes
to surface topography in ventricular myocytes from failing human and rat heart. Proc Natl Acad Sci U S A 2009;
106(16):6854-6859.
(3) Nikolaev VO, Bunemann M, Hein L, Hannawacker A, Lohse MJ. Novel single chain cAMP sensors for receptor-induced signal
propagation. J Biol Chem 2004; 279(36):37215-37218.
(4) Nikolaev VO, Bunemann M, Schmitteckert E, Lohse MJ, Engelhardt S. Cyclic AMP imaging in adult cardiac myocytes reveals
far-reaching beta1-adrenergic but locally confined beta2-adrenergic receptor-mediated signaling. Circ Res 2006;
99(10):1084-1091.
(5) Nikolaev VO, Moshkov A, Lyon AR, Miragoli M, Novak P, Paur H, Lohse MJ, Korchev YE, Harding SE, Gorelik J. {beta}2-Adrenergic
Receptor Redistribution in Heart Failure Changes cAMP Compartmentation. Science. 2010; 26; 327(5973):1653-7.

9

Takeshi Fukuma
Affiliation:
Nano Life Science Institute (WPI-NanoLSI), Kanazawa University

Contact:
E-mail: fukuma@staff.kanazawa-u.ac.jp

Education:
2003 Doctor of Engineering, Department of Electronic Science and Engineering, Kyoto University

Professional Career:
2001-2005

Research Fellow, Kyoto University

2005-2007

Senior Scientist, Physics Department, Trinity College Dublin (Ireland)

2007-2012

Associate Professor, Frontier Science Organization, Kanazawa University

2012-present

Professor, Division of Electronic Eng. and Computer Sci., Professor, Kanazawa University

2017-present

Director/Professor, Nano Life Science Institute (WPI-NanoLSI), Kanazawa University

Scientific Activities:
1999-2003

Instrumentation and applications of atomic-resolution ultrahigh vacuum AFM.

2003-present

Instrumentation and applications of atomic-resolution liquid-environment AFM.

Research Interests: Atomic Force Microscopy, Nanotechnology, Solid-Liquid Interfaces, Nanobio-science
Honors:2008

The 4th Incentive Award for Young Scientist, Biophysical Society of Japan

2010

19th MRS-J Academic Symposium Incentive Award, Materials Research Society of Japan2011

The Young Scientists’Prize, The Commendation for Science and Technology by the MEXT

2017

Hokkoku Bunka Award, Hokkoku Shinbun

Publications:
1.

K. Honbo, S. Ogata, T. Kitagawa, T. Okamoto, N. Kobayashi, I. Sugimoto, S. Shima, A. Fukunaga, C. Takatoh, T. Fukuma, ACS
Nano,10 (2016) 2575-2583.

2.

K. Miyata, J. Tracey, K. Miyazawa, V. Haapasilta, P. Spijker, Y. Kawagoe, A. S. Foster, K. Tsukamoto, T. Fukuma, Nano Lett., 17(2017) 4083-4089.

10Three-Dimensional Atomic Force Microscopy for Visualizing Distribution of Water
and Molecular Chains at Solid-Liquid Interfaces
Takeshi Fukuma
Nano Life Science Institute (WPI-NanoLSI), Kanazawa University
At a solid-liquid interface, water molecules interact with a surface to present non-uniform three-dimensional (3D) distribution
referred to as hydration structures. In addition, atoms and molecules constituting the surface are not necessarily static but
dynamically fluctuating or diffusing to induce various interfacial phenomena. Consequently, a solid-liquid interface is not
two-dimensional but has a three-dimensional extent. However, such 3D fluctuating structures are very difficult to image, which has
limited our understanding on various interfacial phenomena in biology, physical chemistry, electrochemistry, tribology, mineralogy
and environmental sciences. To overcome this problem, we have been working on the development and applications of 3D atomic
force microscopy (3D-AFM) [1]. In this technique, AFM tip is scanned in a 3D interfacial space and the force applied to the tip during
the scan is recorded to produce a 3D force image. During the tip scan, the tip interacts with surrounding water and other flexible
molecules so that the obtained force map represents their time-averaged distribution. So far, this method is mostly used forvisualizing hydration structures on various minerals such as mica, calcite, fluorite and graphite. However, it is also shown that it canbe used for visualizing hydration structures of softer materials such as biological molecules (e.g., bRs, GroELs and DNAs). One ofthese measurements, imaging of a lipid/water interface [2], suggests that the obtained force image reflects not only the hydration
structures but also the 3D distribution of flexible molecular chains. This implies that 3D-AFM may allow us to visualize various 3D
structures as long as the tip can penetrate into the subsurface area without causing irreversible damages. To explore this possibility,
we have been working on visualization of various interfacial structures with different level of complexity and inhomogeneity.

References:[1] T. Fukuma, Y. Ueda, S. Yoshioka, H. Asakawa, “Atomic-Scale Distributionof Water Molecules at the Mica-Water Interface Visualized byThree-Dimensional Scanning Force Microscopy”, Phys. Rev. Lett., 104(2010) 016101.
[2] H. Asakawa, S. Yoshioka, K. Nishimura, T. Fukuma, “Spatial Distributionof Lipid Headgroups and Water Molecules at Membrane/Water
Interfaces Visualized by Three-Dimensional Scanning ForceMicroscopy”, ACS Nano, 6 (2012) 9013-9020.
Fig.

1:

3D-AFM

interfaces.

11

imagingof

nanobioAdam S. Foster
Affiliation:
Aalto University, P.O. Box 11100, FI-00076 AALTO, Finland
Nano Life Science Institute (WPI-NanoLSI), Kanazawa University

Contact:
Phone: +358 504333097
E-mail: adam.foster@aalto.fi

Education:
1997

Masters in Theoretical Physics, Newcastle University, UK

2000

PhD in Theoretical Solid State Physics, University College London, UK

Professional Career:2000 – 2004

Postdoctoral researcher, Helsinki University of Technology, Finland

2004 – 2009

Academy of Finland senior research fellow, Helsinki University of Technology, Finland

2009 – 2012

Professor, Tampere University of Technology, Finland

2012 – present

Professor, Aalto University, Finland

2015 – present

Research professor, Kanazawa University, Japan2017 – present

Visiting professor, Graduate School Materials Science in Mainz, Germany

Research Interests:
Surfaces and Interfaces at the Nanoscale group (SIN), internationally competitive simulation group, computational modelling of
nanoscale phenomena, understanding and controlling the atomic processes dominating function at the surfaces and interfaces ofapplication critical materials

Honors: 2009 – Awarded Väisälä prize by Suomalainen Tiedeakatemia
Publications:
1. Elemental Identification by Combining Atomic Force Microscopy and Kelvin Probe Force Microscopy, Fabian Schulz, Juha Ritala,
Ondrej Krejci, Ari Paavo Seitsonen, Adam S. Foster and Peter Liljeroth, ACS Nano 12 (2018) 5274
2. Multiple hetero-atom substitution in a graphene nanoribbon, Shigeki Kawai, Soichiro Nakatsuka, Takuji Hatakeyama, Rémy
Pawlak, Tobias Meier, John Tracey, Ernst Meyer and Adam S. Foster, Sci. Adv. 4 (2018) eaar7181
3. Machine learning hydrogen adsorption on nanoclusters through structural descriptors, Marc Jäger, Eiaki Morooka, Filippo Federici
Canova, and Adam Foster, npj Computational Materials 4 (2018) 37

12

Probing molecular processes at solid-liquid interfaces
Adam S. Foster
Department of Applied Physics, Aalto University, Finland
Nano Life Science Institute (WPI-NanoLSI), Kanazawa University, Japan
Graduate School Materials Science in Mainz, Germany
The development of high-resolution Atomic Force Microscopy (AFM) in liquids heralded an optimism that reliable atomic andmolecular resolution at solid-liquid interfaces would soon be readily available. AFM is already established as an extremely powerfultool for nanoscale imaging of biological systems in solution, but higher resolutions would open the door to atomic scale
understanding of the wide spectrum of processes occurring in solution. Despite several outstanding successes, high-resolution AFM
in liquids has been plagued by challenges in interpretation and environment control. Many previous studies of the interaction ofwater with solid surfaces have assumed a simple interface, but the latest advances show that functional properties are sensitive toits detailed atomic/electronic structure and dynamics.

Figure 1 Snapshot from an ab initio molecular dynamics
simulation of a TiO2-water interface showing atomic positions andelectron density.

Metal-oxide

Water
In this work, we briefly review our general approach forsimulating high resolution AFM on solid-liquid interfaces [1], butthen introduce recent studies that demonstrate how the
combination of simulations and experiments can be used to gain
insight into dynamic processes, including dissolution [2]. However,
the application of these simulation approaches to more complex

systems, particularly large organic assemblies, requires new methodologies. Here, we will outline our approach to machine learn thefundamental behaviour of water at interfaces as a function of local chemical environment and develop a predictive tool for AFM
simulations that remains rapid for any system.

References:[1] T. Fukuma, B. Reischl, N. Kobayashi, P. Spijker, F. F. Canova, K. Miyazawa, and A. S. Foster, Phys. Rev. B 92 (2015) 155412[2] K. Miyata, J. Tracey, K. Miyazawa, V. Haapasilta, P. Spijker, Y. Kawagoe, A. S. Foster, K. Tsukamoto, and T. Fukuma, Nano Lett., 17(2017) 4083

13

Joshua Edel
Affiliation:
Department of Chemistry, Imperial College London

Contact:
E-mail: joshua.edel@imperial.ac.uk

Education:
2004

PhD in Physical Chemistry, Department of Chemistry, Imperial College London

2000

BSc (Hon) in Chemistry, Department of Chemistry, University of British Columbia

Professional Career:
2016 –

Professor of Biosensing and Analytical Sciences, Department of Chemistry, Imperial

2006 – 2016

Lecturer - Reader, Department of Chemistry, Imperial

2005 – 2006

Research Fellow, Rowland Institute at Harvard, Harvard University

2004 –2005

Postdoctoral Research Assistant, School of Applied and Engineering Physics, Cornell University

Research Interests: Single molecule biological screening using electrical and optical methods, Early stage diagnostics (cancer,
neurodegenerative disorders), single cell screening, electrovariable plasmonics, microfluidics

Honors:2017

ERC Consolidator Investigator Award, EU

2015

President’s Prize for Excellence, Imperial College London, UK

2015

ERC Proof of Concept Award, EU

2012 – 2016

ERC Starting Investigator Award, EU

Publications:
1. Paulose et al., Nanoscale Tweezers for Single Cell Biopsies, Nature Nanotechnology (2018, in press)
2. Cadinu et al., Double Barrel Nanopores as a New Tool for Controlling Single-Molecule Transport, Nano letters (2018)
3. Barik et al., Graphene-edge dielectrophoretic tweezers for trapping of biomolecules, Nature Communications (2017).
4. Montelongo et al., Electrotunable Nanoplasmonic Liquid Mirror, Nature Materials (2017).

14

Novel strategies in molecular sensing
Joshua Edel
Department of Chemistry, Imperial College London
Analytical Sensors plays a crucial role in today’s highly demanding exploration and development of new detection strategies.
Whether it be medicine, biochemistry, bioengineering, or analytical chemistry the goals are essentially the same: 1) improvesensitivity, and 2) maximize throughput. In order to address these key challenges, the analytical community has borrowed
technologies and design philosophies which has been used by the semiconductor industry over the past 20 years. By doing so, key
technological advances have been made which include the miniaturization of sensors and signal processing components whichallows for the efficient detection of nanoscale object. One can imagine that by decreasing the dimensions of a sensor to a scale
similar to that of a nanoscale object, the ultimate in sensitivity can potentially be achieved – the detection of single molecules. This
talk highlights novel strategies for performing single molecule screening.

References:
1. P. Cadinu, G. Campolo, S. Pud, W. Yang, J.B. Edel, C. Dekker, A.P. Ivanov, Double Barrel Nanopores as a New Tool for Controlling
Single-Molecule Transport, Nano letters (2018)
2. R. Ren, Y. Zhang, B.P. Nadappuram, B. Akpinar, D. Klenerman, A.P. Ivanov, J.B. Edel, Y. Korchev, Nanopore extended field-effecttransistor for selective single-molecule biosensing, Nature Communications (2017).
3. X. Lin, A.P. Ivanov, J.B. Edel, Selective single molecule nanopore sensing of proteins using DNA aptamer-functionalised gold
nanoparticles, Chemical Science (2017).
4. J.Y.Y Sze, A.P. Ivanov, A.E.G. Cass, J.B. Edel, Single Molecule Multiplexed Nanopore Protein Screening in Human Serum using
Aptamer modified DNA Carriers, Nature Communications (2017).
5. P. Cadinu et al. Single Molecule Trapping and Sensing using Dual Nanopores separated by a Zeptolitre Nanobridge, Nano Letters(2017).
6. K.J. Freedman, L.M. Otto, A.P. Ivanov, A. Barik, S.H. Oh, J.B. Edel, Nanopore Sensing at Ultra-low Concentrations using Single
Molecule Dielectrophoretic Trapping, Nature Communications (2016).

15

Patrick Unwin
Affiliation:
University of Warwick

Contact:
Department of Chemistry, University of Warwick, UK
E-mail: p.r.unwin@warwick.ac.uk

Education:
1985

BSc (Hons) Chemistry, Class I, University of Liverpool, UK

1989

DPhil, Physical Chemistry, University of Oxford, UK

Professional Career:
1988-90

Junior Research Fellow in Physical Science, Balliol College, University of Oxford

1990-91

SERC/NATO Fellow, University of Texas at Austin, USA

1992-96

Lecturer in Physical Chemistry, University of Warwick

1996-98

Senior Lecturer in Physical Chemistry, University of Warwick

1998-present

Professor of Chemistry, University of Warwick

Scientific Activities:
2010-2015

European Research Council Advanced Grant

2016-present

Editorial Committee of Annual Review of Analytical Chemistry (Annual Reviews series)

2014-present

Director EPSRC-Warwick Molecular Analytical Science Centre for Doctoral Training

Research Interests: Development of fast, ultrasensitive, multifunctional probe imaging techniques that provide a new way of“seeing” electrochemistry and interfacial processes in action, to reveal active surface sites directly, unambiguously andquantitatively.

Honors: Charles N. Reilley Award, Society of Electroanalytical Chemistry, USA (2018); International Society ofElectrochemistry-Elsevier Prize for Experimental Electrochemistry (2017); Royal Society-Wolfson Research Merit Award (2017);
Tilden Prize, Royal Society of Chemistry (2012)

Publications:
1. Nanoscale Structure-Dynamics within Electrocatalytic Materials, J. Am. Chem. Soc., 2017, 139, 16813.
2. Surface Charge Visualization at Viable Living Cells, J. Am. Chem. Soc., 2016, 138, 3152.
3. High-Speed Electrochemical Imaging ACS Nano, 2015, 9, 8942.

16

Multifunctional Nanoscale Imaging with Nanopipettes
Patrick Unwin
University of Warwick, UK
Advances in microscopy and micro-spectroscopy have revealed the chemistry and physical structure of surfaces and interfaces asnever before. Yet, a holistic understanding of interfacial processes, from the functioning of living cells to the operation ofelectrocatalysts, requires spatially-resolved information on other properties, such as chemical flux and surface charge. We havesought to develop microscopy methods that can reveal function and topography synchronously and, further, where highly resolved
flux data can be combined with information from other microscopy approaches in a correlative co-location microscopy strategy, toreveal the relationship between structure and function unambiguously [1,2]. We make use of nanopipettes (with dimensions down
to a few 10s nm) containing electrolyte solution and quasi-reference counter electrodes as mobile probes and electrochemical cells
that can be moved quickly and intelligently near to surfaces to gather multiple electrochemical flux signals that are processed byfinite element method modelling [3]. We shall describe how potential control of the tip, novel feedback and probe scanconfigurations, multi-channel nanopipette probes, and detailed understanding of mass transport in nanopipettes [4], opens up
major new capability of nanopipette imaging probes, with applications ranging from the uptake and delivery of molecules to single
cells [5], surface charge at cell membranes [6] and other surfaces [7], surface patterning and printing [8], and identifying key active
sites on electrode surfaces and electromaterials [9,10], leading to the rational design of electrocatalysts.
I am grateful to many talented students, postdocs, and research fellows at Warwick, and collaborators elsewhere, for their huge
contributions to instrumentation development and applications. I thank the Royal Society for a Wolfson Research Merit Award, andthe ERC and EU for funding which enabled many of the developments herein.
Figure 1. a) Surface charge map of a root hair cell showing enhanced charge magnitude at the tip. b) Map of heterogeneous uptake
rates of a molecule at a root hair cell. c) Electric field profile from FEM simulations at end of a nanopipette near a surface.

References:[1] M. Kang, D. Momotenko, A. Page, D. Perry, and P. R. Unwin, Langmuir, 2016, 32, 7993–8008.
[2] A. Page, D. Perry and P. R. Unwin, Multifunctional Scanning Ion Conductance Microscopy (SICM), Proc. R. Soc. A, 2017, A473
(2200), 20160889.
[3] M. E. Snowden, A. G. Güell, S. C. S. Lai, K. McKelvey, N. Ebejer, M. A. O'Connell, A. W. Colburn and P. R. Unwin, Anal. Chem., 2012,
84, 2483–2491.
[4] D Perry, A Page, B Chen, BG Frenguelli, PR Unwin, Anal. Chem., 2017, 89, 12458–12465.
[5] A. Page, M. Kang, A. Armitstead, D. Perry and P. R. Unwin, Anal. Chem., 2017, 89, 3021–3028.

17[6] D. Perry, B. Paulose Nadappuram, D. Momotenko, P. D. Voyias, A. Page, G. Tripathi, B. G. Frenguelli, and P. R. Unwin, J. Am. Chem.
Soc., 2016, 138, 3152–3160.
[7] D. Perry, R. Al Botros, D. Momotenko, S. L. Kinnear and P. R. Unwin, ACS Nano, 2015, 9, 7266–7276.
[8] D. Momotenko, A. Page, M. Adobes-Vidal and P. R. Unwin, ACS Nano, 2016, 10, 8871–8878.
[9] J. C. Byers , A. G. Güell, P R. Unwin, J. Am. Chem. Soc., 2014, 136, 11252–11255.
[10] C. L. Bentley, M. Kang, and P. R. Unwin, J. Am. Chem. Soc., 2017, 139, 16813–16821.

18

Atsushi Hirao
Affiliation:
Division of Molecular Genetics, Nano Life Science Institute (WPI-NanoLSI)
Cancer Research Institute, Kanazawa University

Contact:
E-mail: ahirao@staff.kanazawa-u.ac.jp

Education:
1988

M.D. Jichi Medical University

1994

Ph.D. University of Tokushima

Professional Career:
1995 - 1997

Postdoctoral fellow, Japan Society for the Promotion of Science (Kumamoto University)

1997 - 2001

Postdoctoral fellow, Ontario Cancer Institute, University of Toronto, Canada

2001 – 2002

Assistant professor, Institute of Molecular Embryology and Genetics, Kumamoto University

2002 - 2004

Assistant professor, Keio University School of Medicine

2004 - 2005

Associate professor, Keio University School of Medicine

2005 – present

Professor, Cancer Research Institute, Kanazawa University

2017- present

Director, Cancer Research Institute, Kanazawa University

Scientific Activities:
2011-present

Associate editor of Cancer Science

2014-present

Associate member of Science Council of Japan2017-present

Executive director of Japanese Society of Hematology

Research Interests: Stem cell biology, Metabolism, Cancer therapy
Honors:2007

The JSPS (Japan Society for the Promotion of Science) PRIZE

2010

The Commendation for Science and Technology by the MEXT

Publications:
1. Tadokoro Y, et.al., Cell Stem Cell. 22:713-725, 2018
2. Hoshii T, et.al., Proc Natl Acad Sci U S A. 111:3805-10, 2014
3. Naka K, et.al., Nature, 463:676-80, 2010
4. Miyamoto K, et.al., Cell Stem Cell. 1:101-112, 2007

19

Metabolic control of cancer stemness
Atsushi Hirao, MD, PhD
Division of Molecular Genetics, Nano Life Science Institute (WPI-Nano LSI),
Cancer Research Institute, Kanazawa University
Nutrients, such as amino acids and glucose, are critical determinants of cell survival, proliferation and differentiation processes innormal and malignant tissues. Recent studies have revealed critical roles of metabolic control in stem cell properties, so called"stemness"，which contribute to malignant progression of cancers. It has been shown that metabolic control is critical for
maintenance of hematopoietic stem cell (HSCs) functions. HSCs utilize glycolysis to sustain undifferentiated status, whereas
oxidative phosphorylation becomes dominant in progenitors. Unlike HSCs, leukemia stem cells (LSCs) appear to have low glycolytic
flow, indicating that there are critical differences in metabolic regulation between these cell types. Therefore, knowledge of themolecular mechanisms of metabolic regulation will provide novel therapeutic approaches for eradiation of LSCs. We previously
found that fine-tuning of mTOR complex 1 (mTORC1) and forkhead transcription factor FoxO, which function in nutrient sensing
signaling pathways, is critical for maintenance of HSC homepostasis. We found that modification of these signals induces apoptosis
of LSCs, but not HSCs. Therefore, we believe that investigation of molecular functions in these pathways will lead to thedevelopment of successful therapeutics for eradiation of LSCs.
In the WPI program, we aim to understand the molecular mechanisms of metabolic regulation for determining stem cell fate innormal and malignant tissues by using Bio-SPM. To do so, we will identify metabolic enzymes that play critical role in cancer
malignancy by CRISPR library as therapeutic targets, followed by identification of metabolites as biomarkers. We also develop novel
probes that specifically bind to particular metabolites in collaboration with experts of supermolecular chemistry. Finally, we will
combine the probe with SPM, such as SICM. By collaborations with experts in nanotechnology, we shall build a path to a new era ofcancer biology.

References:[1] Tadokoro Y, et.al., Spred1 Safeguards Hematopoietic Homeostasis against Diet-Induced Systemic Stress. Cell Stem Cell, 2018,
22:713-725.
[2] Hegazy AM, et al., Therapeutic strategy for targeting aggressive malignant gliomas by disrupting their energy balance. J Biol
Chem., 2016, 291:21496-21509.
[3] Hoshii, T., et al., mTORC1 is essential for leukemia propagation but not stem cell self-renewal. J. Clin. Invest., 2012, 122:
2114-2129.
[4] Naka K, et al., TGF -FOXO signalling maintains leukaemia-initiating cells in chronic myeoid leukaemia. Nature, 2010, 463:676-80.
[5] Miyamoto K, et al., Foxo3a is essential for maintenance of the hematopoietic stem cell pool. Cell Stem Cell, 2007, 1:101-12.

20

Mark MacLachlan
Affiliation:
Dept. of Chemistry, University of British Columbia
Nano Life Science Institute (WPI-NanoLSI), Kanazawa University

Contact:
2036 Main Mall, Vancouver, BC, Canada V6T 1Z1
E-mail: mmaclach@chem.ubc.ca

Education:
1995

B.Sc.(Hons.) Chemistry, University of British Columbia

1999

Ph.D. Chemistry, University of Toronto

Professional Career:
1999-2001

NSERC PDF, M.I.T.

2001-2007

Assistant Professor, UBC

2007-2011

Associate Professor, UBC

2011-present

Professor, UBC

2016-present

Associate Dean of Research & Graduate Studies, UBC

2017-present

Visiting Research Professor, Kanazawa University

Scientific Activities:
2014-present

Fellow, Royal Society of Canada

2015-present

Director, NSERC CREATE Training Prog. in Nanomaterials Science & Technol. (UBC)

2016-2018

Advisory Board Member, Materials Chemistry Frontiers, Royal Society of Chemistry

Research Interests: Supramolecular chemistry, macrocycles, nanomaterials, cellulose nanocrystals, porous materials.
Honors:Strem Award for Pure or Applied Inorganic Chemistry (Canadian Society for Chemistry (CSC), 2013), Rutherford Medal (Royal Societyof Canada (RSC), 2013), Invitational Fellowship for Research in Japan (JSPS, 2013), Steacie Prize (2014), Award for Excellence inMaterials Chemistry (CSC, 2016)

Publications:
1. Wang, P. X.; Hamad, W. Y.; MacLachlan, M. J. Angew. Chem. Int. Ed. 2018, 57, 3360-3365.
2. Wang, P.-X.; Hamad, W. Y.; MacLachlan, M. J. Nature Comm. 2016, 7, 11515.
3. Shopsowitz, K.E.; Qi, H.; Hamad, W.Y.; MacLachlan, M.J. Nature 2010, 468, 422-425.

21

Cellulose Nanocrystals: Materials for Photonic Structures
Mark MacLachlan
Dept. of Chemistry, University of British Columbia
Nano Life Science Institute (WPI-NanoLSI), Kanazawa University
Cellulose nanocrystals (CNCs) can be extracted from biomass, including cotton and wood pulp. When prepared throughsulfuric acid-catalyzed hydrolysis of biomass, CNCs are obtained with sulfate ester groups on their surface. The
resulting CNCs (Fig. 1) form stable colloidal suspensions in water, but they also organize into a chiral nematic liquid
crystal when concentrated above ~4 wt% in water. This chiral nematic organization is retained in dried films of CNCs,
giving films with bright, iridescent colors.[1]Our group has been active in using CNCs as a template to construct novel chiral nematic mesoporous materials withphotonic properties.[2] We have succeeded in preparing silica,[3] organosilica,[4] and polymeric materials[5] throughself-assembly or through hard-templating methods.
In this presentation, I will introduce nanoscale biomaterials, illustrate our concept for creating new materials withtunable photonic properties (such as the organosilica glass shown in Fig. 1), and highlight some of our recent work inthis area.[6-8]

Figure 1 TEM image of CNCs (left) and photograph of chiral nematic, mesoporous organosilica (right).

References:[1] J.-F. Revol, L. Godbout and D. G. Gray, J. Pulp Pap. Sci., 1998, 24, 146.
[2] M. Giese, L .K. Blusch, M. K. Khan and M. J. MacLachlan, Angew. Chem. Int. Ed., 2015, 54, 2888.
[3] K. E. Shopsowitz, H. Qi, W. Y. Hamad and M. J. MacLachlan, Nature, 2010, 468, 422.
[4] K. E. Shopsowitz, W. Y. Hamad and M. J. MacLachlan, J. Am. Chem. Soc., 2012, 134, 867.
[5] M. K. Khan, A. Bsoul, K. Walus, W. Y. Hamad and M. J. MacLachlan, Angew. Chem. Int. Ed., 2015, 54, 4304.
[6] A. Tran, W. Y. Hamad and M. J. MacLachlan, ACS Appl. Nano Materials, 2018, 1, 3098.
[7] T. Hiratani, O. Kose, W. Y. Hamad and M. J. MacLachlan, Mater. Horizons, 2018, in press.
[8] Y. Cao, W. Y. Hamad and M. J. MacLachlan, Adv. Opt. Mater., 2018, 6, 1800412.

22

David Klenerman
Affiliation:
University of Cambridge

Contact:
E-mail: dk10012@cam.ac.uk

Education:
1979-1982

BA Chemistry, University of Cambridge

1982-1985

PhD Chemistry, University of Cambridge, Department of Chemistry(Supervisor: Professor Ian Smith FRS)

1985-1987

Fulbright Scholar, Stanford University, California (Supervisor: Professor Richard N. Zare)

Professional Career:
1987-1994

BP Research Centre, Sunbury – Research Scientist, Laser Spectroscopy Group

1994-1999

Assistant Director of Research, University of Cambridge

1999-2004

Lecturer, Department of Chemistry, University of Cambridge

2002-2007

Reader in Biophysical Chemistry, University of Cambridge

2007-Present

Professor of Biophysical Chemistry, University of Cambridge

Research Interests: Single molecule fluorescence, scanning ion conductance microscopy, molecular basis of neurodegenerative
disease, molecular basis of T-cell triggering.

Honors:2012

Elected Fellow of the Royal Society (FRS)2015

Elected Fellow of the Academy of Medical Sciences (FMedSci)

2016

Royal Society Glaxo Wellcome Professor of Molecular Medicine

Publications:
1. Initiation of T cell signaling by CD45 segregation at 'close contacts'. Nature Immunology 2016, 17, 574.
2. Direct Observation of the Interconversion of Normal and Toxic Forms of alpha-Synuclein. Cell 2012, 149, 1048.
3. Inhibiting the Ca2+ Influx Induced by Human CSF. Cell Rep 2017 21, 3310.

23

New fluorescence based methods for imaging single molecules
David Klenerman
University of Cambridge
A major advance in physical chemistry in the last two decades has been the development of quantitative methods to directly
observe individual molecules in solution, attached to surfaces, in the membrane of live cells or more recently inside live cells. Theseadvances allow the study of how the individual cellular components interact to form a living cell and how this can go awry in disease.
My group has focused on developing these methods and then, with our biological and clinical collaborators, applying them tobiological and biomedical problems which have been difficult or impossible to solve using more conventional bulk methods. I willshow how we have developed single molecule fluorescence to detect the interaction of molecules in the test-tube and on cells andcombined this with pipette based delivery. I will then show how these methods can be used to gain new molecular insights bydescribing our recent work on the role of proteins aggregates in neurodegenerative disease and the molecular basis of the immune
response.

24

Bart Hoogenboom
Affiliation:
London Centre for Nanotechnology, University College London

Contact:
17-19 Gordon Street, London WC1H 0AH
E-mail: b.hoogenboom@ucl.ac.uk

Education:
1998

MSc in Physics, University of Groningen (NL)2002

PhD in Condensed Matter Physics, University of Geneva (CH)

Professional Career:
2002-2004

Postdoc at the Department of Physics, University of Basel (CH)2005-2007

Postdoc at the Biozentrum, University of Basel (CH)2007-2013

Lecturer (UK eq. Assistant Professor), University College London

2013-2016

Reader (UK eq. Associate Professor), University College London

2016-Present

Professor of Biophysics, University College London

Scientific Activities:
1998-2002

Correlated electron systems, superconductivity, STM/STS

2005-Present

Nanoscale biophysics

Research Interests:
In my lab, we are interested in the broad question of how biological molecules interact which each other and (self-)organise tocollectively carry out tasks that are important for health and disease. Examples of such tasks are the repair of DNA damage, theperforation of cellular membranes and the regulation of transport into and out of the cell nucleus. While we use a range ofbiophysical techniques, our favourite tool for this is atomic force microscopy (AFM).

Honors:Medal for Scanning Probe Microscopy 2017, of the Royal Microscopical Society

Publications:
1.

C. Leung, A. W. Hodel, A. J. Brennan, N. Lukoyanova, S. Tran, C. House, S. C. Kondos, J. C. Whisstock, M. A. Dunstone, J. A. Trapani, I. Voskoboinik,
H. R. Saibil, and B. W. Hoogenboom, Real-time visualization of perforin nanopore assembly, Nature Nanotechnology 12 (2017): 467–473.

2.

A. Bestembayeva, A. Kramer, A. A. Labokha, D. Osmanovic, I. Liashkovich, E. V. Orlova, I. J. Ford, G. Charras, A. Fassati, and B. W. Hoogenboom,
Nanoscale stiffness topography reveals structure and mechanics of the transport barrier in intact nuclear pore complexes, Nature
Nanotechnology 10 (2015): 60–64.

25Assembly and membrane perforation by immune proteinsBart Hoogenboom

London Centre for Nanotechnology, University College London
Pore-forming proteins are crucial armaments in the continuous battle between organisms and their pathogens. In the course of theirfunction, they assemble into oligomeric rings that puncture holes through target membranes. Bacterial toxins are pore-forming
proteins [1,2] that penetrate host cell membranes to release nutrients for the bacteria, thus invading or killing the host cells, forexample in bacterial pneumonia or meningitis. On the other hand, the immune system of vertebrates secretes pore-forming
proteins to kill infected or cancerous cells or invading pathogens. To perform these tasks, pore-forming proteins convert fromwater-soluble monomers into membrane-bound, oligomeric pores. I will report on experiments in which we disentangle thedifferent steps of this intricate process, largely by real-time and nanometer-scale visualisation, focusing on proteins that play key
roles in our immune system [3-5].

Fig. 1. Schematic of perforin nanopore assembly in the immunological synapse between a killer lymphocyte and a transformed orvirus-infected cell [3]. Courtesy of Adrian Hodel.

References:
1.

C. Leung, N. V. Dudkina, N. Lukoyanova, A. W. Hodel, I. Farabella, A. P. Pandurangan, N. Jahan, M. P. Damaso, D. Osmanovic, C. F.
Reboul, M. A. Dunstone, P. W. Andrew, R. Lonnen, M. Topf, H. R. Saibil, and B. W. Hoogenboom, Stepwise visualization of
membrane pore formation by suilysin, a bacterial cholesterol-dependent cytolysin, eLife 3 (2014): e04247.

2.

Adrian W Hodel, Carl Leung, Natalya V Dudkina, Helen R Saibil, and Bart W Hoogenboom, Atomic force microscopy ofmembrane pore formation by cholesterol dependent cytolysins, Current Opinion in Structural Biology 39 (2016): 8–15.

3.

C. Leung, A. W. Hodel, A. J. Brennan, N. Lukoyanova, S. Tran, C. House, S. C. Kondos, J. C. Whisstock, M. A. Dunstone, J. A.
Trapani, I. Voskoboinik, H. R. Saibil, and B. W. Hoogenboom, Real-time visualization of perforin nanopore assembly, Nature
Nanotechnology 12 (2017): 467–473.

4.

D. A. C. Heesterbeek, B. W. Bardoel, E. S. Parsons, I. Bennett, M. Ruyken, D. J. Doorduijn, R. D. Gorham Jr., E. T. M. Berends, A. L.
B. Pyne, B. W. Hoogenboom, and S. H. M. Rooijakkers, Bacterial killing by complement requires membrane attack complex
formation via surface-bound C5 convertases, under revision for EMBO Journal.

5.

E. S. Parsons et al., Pathway and kinetics of assembly of a hetero-oligomeric immune pore, in preparation.

26

Richard Wong
Affiliation:
Nano Life Science Institute (WPI-NanoLSI), Kanazawa University
Institute for Frontier Science Initiative, Kanazawa University

Contact:
E-mail: rwong@staff.kanazawa-u.ac.jp

Education:
2000

M. Phil. (Pediatrics) The University of Hong Kong

2004

Ph.D. (Anatomy & Cell Biology) The University of Tokyo

Professional Career:
2004-2005

JSPS Postdoctoral fellow, Department of Anatomy & Cell Biology, The University of Tokyo

2005-2007

HHMI Research Associate, Laboratory of Cell Biology, The Rockefeller University (USA)

2008-2012

Associate Professor (Tenure-Track), Frontier Science Organization, Kanazawa University

2012-Present

Professor, Biology Course, Faculty of Natural System, Kanazawa University

2015-2018

Professor & Head of Division in Molecule & Cell Research, Bio-AFM Frontier Research Center, Kanazawa
University

2015-Present

Professor & unit leader in Institute for Frontier Science Initiative, Kanazawa University

2017-Present

Professor & Principle investigator in Nano Life Science Institute(WPI-NanoLSI), Kanazawa University

Scientific Activities:
2015-

Referee for Fonds de la Recherche Scientifique, Belgium.

2015-

Referee for Fondation pour la Recherche Médicale, France.

2008- 2018

Visiting Professorship, Laboratory of Cell Biology, The Rockefeller University, USA.

2013-

Visiting Professorship, Department of Hematology, West China Hospital, Sichuan University

Research Interests:
To elucidate the structure and function of Nuclear Pore Complex (NPC) in cells

Honors:2017-

Fellow, The Royal Society of Medicine, United Kingdom.

2017-

Ambassador, American Society of Cell Biology, USA.

Publications:
1. Lim KS and Wong RW Cell Chem. Biol. 2018 25(9):1056-1058.

27

The nuclear pore complex: nanomachine as gatekeeper of molecular transport
across nuclear membranes
Richard Wong

Nano Life Science Institute (WPI-NanoLSI); Institute for Frontier Science Initiative; School of Natural
System, Institute of Science and Engineering; Bio-AFM Frontier Research Center; Kanazawa
University, Kanazawa, Ishikawa, JapanA cell is like a country. The central government is the nucleus. On the surface of the nucleus, there are thousands of tiny turnstiles
and gates (nuclear pores) that are embedded into nuclear membranes. These gates are multiprotein nuclear pore complexes (NPCs).
NPCs are the macromolecular turnstiles between the cytoplasm and the nucleus that control the trafficking of proteins, RNAs andviruses. The giant NPC structures are extremely complex [1]. NPCs modulate nuclear shape and organization and have implicated invarious cellular activities, such as transcription, autophagy [2], mitosis [3] and cell fate determination [4]. Small molecules can passfreely through NPCs; in contrast, larger molecules (>40 kDa) can pass through the pore smoothly only if they are bound to specifictransporter proteins that interact with FG (phenylalanine-glycine)-nucleoporins (FG-Nups), which form a soft and flexible lining
composed of approximately 200 intrinsically disordered polypeptide chains inside the turnstile. Several FG-Nup trafficking models
have been proposed, such as forest, virtual gating, polymer brush, and selective phase/hydrogel models. Unfortunately, there has
been a lack of techniques that can visualize and investigate FG-Nup behavior inside the NPCs of cancer cells. Recently, we show thathigh-speed atomic force microscopy (HS-AFM) can be used to visualize dynamic changes in nuclear pores in cancer cells [5]. We
showed nanoscopic changes of the symmetric pores by morphometric analysis in colon cancer cell. To determine FG-Nup behavior,
we concentrated on the central turnstile enclosed by the cytoplasmic filaments. We visualized that FG-Nups are short, stiff, hair-like,
twisted ropes having a brush-like manner or forming a broken cobweb shape from their tethering point. We propose a“spiders-spidermen working inside the sticky cobweb” model for the process of central channel shuttling or gene gating by the
nuclear pore [5].

References:[1] R.W. Wong Cell Chem. Biol. 2015, 22, 1285-1287.
[2] D.J. Klionsky, K. Abdelmohsen, A. Abe, ... R.W. Wong et al. Autophagy 2016, 12, 1-222. (*Web of Science Citation Index >1200)[3] F.R.P. Dewi, T. Domoto, M. Hazawa, A. Kobayashi, T. Douwaki, T. Minamoto and R.W. Wong

Oncotarget 2018, 9, 13337-13352.

[4] M. Hazawa, D. Lin, A. Kobayashi, Y.Y. Jiang, F.R.P. Dewi, M.S. Mohamed, Hartono, S. Nakada, M. Meguro-Horike, S. Horike, H.
Koeffler and R.W. Wong. EMBO Rep. 2018, 19, 73-88.
[5] M.S. Mohamed, A. Kobayashi, A. Taoka, T. Watanabe-Nakayama, Y. Kikuchi, M. Hazawa, T. Minamoto, Y. Fukumori, N. Kodera, T.
Uchihashi, T. Ando, R.W. Wong. ACS Nano. 2017, 11, 5567-5578.

28

Kunio Matsumoto
Affiliation:
Nano Life Science Institute (WPI-NanoLSI),
Cancer Research Institute, Kanazawa University

Contact:
E-mail: kmatsu@staff.kanazawa-u.ac.jp

Education:
1981

Faculty of Science, Kanazawa University

1983

Graduate School of Science, Kanazawa University

1986

Graduate School of Science, Osaka University (Ph.D.)

Professional Career:
1987-1990

Assistant Professor, Department of Dermatology, Osaka University

1990-1993

Assistant Professor, Department of Biology, Kyushu University

1993-1994

Assistant Professor, Biomedical Research Center, Osaka University

1994-2007

Associate Professor, Graduate School of Medicine, Osaka University

2007-present

Professor, Cancer Research Institute, Kanazawa University

2017-present

Professor, Nano Life Science Institute (WPI-NanoLSI), Kanazawa University

Honors:1992

Academic Incitement Award of Japanese Industrial Hygiene Dermatology Association

1996

JB Award of the Japanese Biochemical Society

1997

Incitement Award of the Japanese Cancer Association
The 3rd Bio Business Competition JAPAN, The Special Award
Nature Medicine-AnGes MG BioMedical Award (Main Award)

2014

The Commendation for Science and Technology by MEXT, Prizes for Science and Technology

Publications:
1.

MiaoW, Sakai K, Imamura R, Ito K, Suga H, Sakuma T, Yamamoto T, Matsumoto K. MET activation by a macrocyclic peptide
agonist couples to biological responses differently from HGF in a context-dependent manner. Int J Mol Sci,19: 3141, 2018.

2.

Imamura R, Matsumoto K. Hepatocyte growth factor in physiology and infectious diseases. Cytokine, 98: 97-106, 2017.

3.

Adachi E, Sakai K, Nishiuchi T, Imamura R, Sato H, Matsumoto K. Cell-autonomous changes in Met receptor expression regulate
the growth and metastatic characteristics in malignant melanoma. Oncotarget, 7: 70779-70793, 2016.

4.

Isozaki H, Ichihara E, Takigawa N, Ohashi K, Ochi N, Yasugi M, Ninomiya T, Yamane H, Hotta K, Sakai K, Matsumoto K, Hosokawa S,
Bessho A, Sendo T, Tanimoto M, Kiura K. Non-small cell lung cancer cells acquire resistance to the ALK inhibitor alectinib byactivating alternative receptor tyrosine kinases. Cancer Res, 76: 1506-1516, 2016.

29

Macrocyclic Peptide Technology Targeting HGF and MET receptor
Kunio Matsumoto

Nano Life Science Institute (WPI-NanoLSI), Cancer Research Institute, Kanazawa University
Hepatocyte growth factor (HGF) is bioactive protein composed of 697 amino acids. HGF binds to and activates the METtransmembrane receptor. HGF-induced activation of the MET receptor drives mitogenesis (cell proliferation), motogenesis (cellmigration), morphogenesis (3-D tubulogenesis), and cell survival, thereby participates in regeneration and protection of tissues,
including the liver and nervous tissue. Clinical trials using recombinant HGF protein are ongoing for treatment of spinal cord injury
and amyotrophic lateral sclerosis. In cancer tissues, aberrant activation of MET receptor by amplification of MET gene and/or
overexpression of HGF participate in malignant progression of cancer such as drug resistance and invasion-metastasis.

Using RaPID (Random Peptide Integrated Discovery) system, an innovative drug discovery platform, we obtained macrocyclic
peptides that specifically bind to HGF or MET with 1 - 10 nM Kd values. The cross-linking of MET-binding macrocyclic peptides,
thereby capable of bivalent display of MET-binding peptides, conferred them an ability to selectively activate MET [1, 2]. These
artificial MET-ligands induced dimerization and activation of
MET receptor, and exhibited mitogenic, motogenic, andmorphogenic activities that are comparable to native ligand
HGF. Our approach for generating bivalent macrocyclic
peptides as non-protein ligands for cell surface receptors
may be useful for developing surrogate ligands with a broad
range of potential applications.

HGF is secreted extracellularly as biologically inactive
single-chain HGF (scHGF). Protease-medicated processing
into two-chain HGF (tcHGF) is critical step for activation of
HGF. In normal tissues, HGF exists as inactive scHGF, while tcHGF is generated in restricted tissues such as cancer tissues. The
selective detection and inhibition of tcHGF are important for cancer diagnosis and therapeutics.

References:[1] Ito K, Sakai K, Suzuki Y, Ozawa N, Hatta T, Natsume T, Matsumoto K, Suga H. Artificial human Met agonists based on macrocycle
scaffolds. Nat Commun, 6: 6373, 2015.
[2] MiaoW, Sakai K, Imamura R, Ito K, Suga H, Sakuma T, Yamamoto T, Matsumoto K. MET activation by a macrocyclic peptide agonist
couples to biological responses differently from HGF in a context-dependent manner. Int J Mol Sci,19: 3141, 2018.

30