From Molecules to Cells and Tissue– Bridging Scaleswith Nanoprobe Technology

7th
NanoLSI
Symposiumin Berlin
NOVEMBER

2-3, 2023

7th
NanoLSI
Symposium
From Molecules to Cells and Tissue– Bridging Scaleswith Nanoprobe Technology

NOVEMBER 2-3, 2023
VENUE
HARNACK HOUSE THE CONFERENCE VENUE OF THE MAX PLANCK SOCIETY
IHNESTR. 1620  14195 BERLIN, GERMANY

01

Welcome Message fromthe Organizer and Co-Organizerof the 7th NanoLSI Symposium
It is a great pleasure to welcome you to the 7th NanoLSI Symposium in Berlin. The Nano Life
Science Institute (WPI-NanoLSI) at Kanazawa University aims at a fundamental
understanding of a wide range of life-related phenomena through transdisciplinary research in
the fields of Nanometrology, Life Science, Supramolecular Chemistry, and Computational
Science. At this international symposium, distinguished researchers from Japan, Canada,
Switzerland, and Germany will gather for two days to promote interdisciplinary research andnew collaborations among scientists from diverse backgrounds, and to discuss cutting-edge
nanoprobe technologies and their impact across a wide range of scales: “From Molecules to
Cells and Tissue – Bridging Scales with Nanoprobe Technology”. We are delighted towelcome all participants to our historic venue at Harnack House and we are looking forward
to interesting and stimulating presentations and discussions.

c Kirsten Sachse

02

Carsten Beta

Clemens M. Franz

Organizer

Co-Organizer

Carsten Beta

Clemens M. Franz

Professor of Biological Physics at the
University of Potsdam, Germany, andNanoLSI Overseas PI at Kanazawa
University, Japan — works at the interface ofphysics and biology. His researchconcentrates on cell motility and chemotaxis
of both bacterial swimmers and amoeboid
cells and is based on experimental studies
using live-cell imaging and microfluidics, aswell as theoretical modeling inspired byconcepts of active matter physics, nonlinear
dynamics, and pattern formation.

Associate Professor at NanoLSI, Kanazawa
University, Japan – applies atomic force
microscopy (AFM) and other advanced
microscopy techniques to study cell adhesion
and mechanotransduction mechanisms.
His research focuses on quantitating
receptor-mediated adhesion forces,
visualizing dynamic receptor-ligand
complexes by high-speed AFM, as well as
intracellular exploration by large-scale
scanning probe microscopy.

03

7th NanoLSI Symposium
November 2, 2023 / Day 1

8:50 AM – 6:30 PM (CET)(Open venue: 8:00 AM)

November 3, 2023 / Day 2

8:00 AM

Registration and arrival tea & coffee

8:30 AM

Registration and arrival tea & coffee

8:50 AM

Opening remarks: Takeshi Fukuma, Director of Nano Life Science Institute, Kanazawa University

9:00 AM

Poster session

9:00 AM

Session #1 9:00 AM – 11:00 AM
Chairpersons:Session theme:

10:00 AM

Clemens Franz & Noriyuki Kodera
Proteins in operation(Open venue: 8:30 AM)@Meitner Hall

Session #4

10:00 AM – 12:00 PM

Chairperson:
Session theme:

Masanobu Oshima
Single cell dynamics

1. Noriyuki Kodera (NanoLSI, Kanazawa University)
Video imaging of dynamic behaviors of biomolecules by high-speed atomic force microscopy

1. Takeshi Fukuma (NanoLSI, Kanazawa University)
Visualizing nanoscale dynamics and mechanics in living cells by nanoendoscopy AFM

2. Ulrich Schwarz (Heidelberg University)
Adsorption and self-assembly of SAS-6 rings on a surface studied with high-speed AFM and computer simulations

2. Salvatore Chiantia (University of Potsdam)
Investigating virus-host interactions at a single cell level using quantitative fluorescence microscopy

3. Clemens Franz (NanoLSI, Kanazawa University)
Investigating actomyosin contractility-driven adhesion modulation by complementary high-speed AFM andfluorescence microscopy

3. Daniel Müller (ETH Zürich)Mechanically quantifying and guiding biological processes
4. Tomoyoshi Yamano (Graduate School of Medical Sciences / NanoLSI, Kanazawa University)
Development of engineered exosome for immune regulation

4. Erik Schäffer (University of Tübingen)Ultraresolution picotensiometry of kinesin motors

11:00 AM

9:00 AM – 1:30 PM (CET)

Poster session @Meitner Hall

12:00 PM

Closing remarks

1:00 PM

Lunch

12:30 PM

Lunch 12:30 PM – 1:30 PM

2:00 PM

Session #2

Carsten Beta, Organizer of the 7th NanoLSI Symposium, University of Potsdam/NanoLSI

2:00 PM – 4:00 PM

Chairperson: Takeshi Fukuma
Session theme: From cells to tissue
1. Masanobu Oshima (NanoLSI/CRI, Kanazawa University)
Identification of genotype-linked nano-scale physical properties of intestinal tumor cells
2. Anna Taubenberger (Technische Universität Dresden)
Quantifying tumor spheroid mechanical properties in confining 3D microenvironments
3. Satoru Okuda (NanoLSI, Kanazawa University)
Versatile computational modeling of 3D multicellular dynamics and application to collective cell migration
4. Martin Falcke (Max Delbrück Center for Molecular Medicine)
On multistability and constitutive relations of cell motion on fibronectin lanes

4:00 PMTea & coffee break

4:30 PM

Session #3

4:30 PM – 6:30 PM

Chairpersons:Session theme:

Mark MacLachlan & Satoru Okuda
Material properties and control
1. Mark MacLachlan (The University of British Columbia/NanoLSI, Kanazawa University)
Supramolecular chemistry using bionanomaterials
2. Martin Bastmeyer (Karlsruhe Institute of Technology)
3D cellular microenvironments to study cell mechanics
3. Svetlana Santer (University of Potsdam)
From molecular actuation to microscopic motion
4. Shinji Watanabe (NanoLSI, Kanazawa University)
Time-resolved imaging of nanomechanics and nanostructures of living cells with scanning ion conductance microscopy

04

6:30 PM

Photo session

7:00 PM

Dinner buffet (by invivation only)

7:00 PM – 9:30 PM

05

ORAL SESSIONS

06

07

Noriyuki Kodera

https://researchmap.jp/
NoriyukiKodera?lang=en

Video Imaging of Dynamic Behaviors of Biomolecules
by High-Speed Atomic Force Microscopy

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

Noriyuki Kodera

E-mail: nkodera@staff.kanazawa-u.ac.jp
Website: https://researchmap.jp/NoriyukiKodera?lang=en

Research Interests
Biophysics, molecular motors, cytoskeletons, atomic force microscopy

Education
2005
2003
2001

Ph.D., Div. of Basic Sci., Grad. Sch. Nat. Sci. & Tech., Kanazawa Univ.
M. Sci., Div. of Math. & Phys. Sci., Grad. Sch. Nat. Sci. & Tech., Kanazawa Univ.
B. Sci., Dept. of Phys, Fac. of Sci., Kanazawa Univ.

Professional Experience
2018 – present2013 – 20172010 – 20182005 – 2010

Prof., WPI-NanoLSI, Kanazawa Univ.
PRESTO Researcher, JST
Asst. Prof. & Assoc. Prof., Bio-AFM Frontier Research Center, Inst. Sci. & Eng., Kanazawa Univ.
Research Fellow (DC2 & PD), JSPS and Postdoctoral Fellow, CREST, JST

Honors
2017
2013
2012

JSPS Prize

High-speed atomic force microscopy (HS-AFM) is a unique microcopy that allows direct real-timevisualization of biological macromolecules in action under near-physiological conditions, without any
chemical labeling. Typically, the temporal resolution is sub-100 ms, and the spatial resolution is 2–3
nm in the lateral direction and ~0.1 nm in the vertical direction [1]. A wide range of biomolecularsystems and their dynamic processes have been studied by HS-AFM, providing deep mechanistic
insights into how biomolecules function. Notably, HS-AFM is particularly powerful in visualizing large
and flexible biomolecular systems, which conventional structural biology methods are unable tovisualize. These unique features of HS-AFM are well demonstrated in recent research results thatvisualize the structural dynamics of intrinsically disordered regions, single polypeptide chains withheight of ~0.5 nm in a protein [2], the gathering of translation factors around the ribosomal stalkcomplex [3], and the long-range DNA degradation of CRISPR-Cas3 driven by its helicase andnuclease activities [4].
In the presentation, after overviewing the principles and performance of HS-AFM, AFM images
showing dynamic behaviors of biomolecules will be shown. In addition, our recent efforts to improve
the speed performance of HS-AFM will be discussed [5].

References
1.
2.

Award for Science and Technology (Development Category), The Commendation for Scienceand Technology by the Minister of Education, Culture, Sports, Science and Technology

3.

Young Researcher’ s Nanoprobe Technology Prize (JSPS Nanoprobe Technology 167)

4.

Publications

08

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

1.

Yoshimi K, Takeshita K, Kodera N, …, Mashimo T, “Dynamic mechanisms of CRISPR interference by Escherichia coli CRISPR-Cas3” .
Nature Communications 13, 4917 (2022).

2.

Kodera N, Noshiro D, Dora SK, …, Longhi S, Ando T, "Structural and dynamics analysis of intrinsically disordered proteins by
high-speed atomic force microscopy". Nature Nanotechnology 16, 181-189 (2021).

3.

Imai H, Uchiumi T, Kodera N, "Direct visualization of translational GTPase factor pool formed around the archaeal ribosomal P-stalk by
high-speed AFM". PNAS 117, 32386-32394 (2020).

5.

Ando, T.; Uchihashi, T.; Kodera, N., High-speed AFM and applications to biomolecular systems. Anuu. Rev. Biophys. 2013, 42, 393-414.
Kodera, N.; Noshiro, D., Dora, S. K.; … ; Longhi, S.; Ando T., Structural and dynamics analysis of intrinsically disordered proteins
by high-speed atomic force microscopy. Nat. Nanotechnol. 2021, 16, 181-189.
Imai, H.; Uchiumi, T.; Kodera, N., Direct visualization of translational GTPase factor pool formed around the archaeal
ribosomal P-stalk by high-speed AFM. PNAS 2020, 117, 32386-32394.
Yoshimi, K.; Takeshita, K.; Kodera, N.; … ; Mashimo, T., Dynamic mechanisms of CRISPR interference by Escherichia coli
CRISPR-Cas3. Nat. Commun. 2022, 13, 4917.
Umeda, K.; McArthur, S. J.; Kodera, N., Spatiotemporal resolution in high-speed atomic force microscopy for studying
biological macromolecules in action. Microscopy 2023, 72, 151-161.

09

Ulrich S. Schwarz

https://www.thphys.uniheidelberg.de/~biophys/

Professor for Theoretical Physics

Adsorption and Self-assemblyof SAS-6 Rings on a Surface Studied
with High-Speed AFM and Computer Simulations

Institute for Theoretical Physics and BioQuant, Heidelberg University, Germany

Ulrich S. Schwarz

E-mail: schwarz@thphys.uni-heidelberg.de
Website: https://www.thphys.uni-heidelberg.de/~biophys/

Research Interests
Theoretical biophysics, cell mechanics and adhesion, cell organization, self-assembly

Education
1998
1994

PhD in theoretical physics MPI and U Potsdam
Physics diploma LMU Munich

Professional Experience2009 – present
2008 – 2009
2001 – 2007
1998 – 2000

Full professor for Theoretical Physics at Heidelberg University
Full professor for Theoretical Biophysics at Karlsruhe Institute of Technology (KIT)
Emmy Noether Junior Research Group Leader at MPI Potsdam and U Heidelberg
Postdoc Weizmann Institute, Israel

Institute for Theoretical Physics and BioQuant, Heidelberg University, Germany

Spindle assembly abnormal protein 6 (SAS-6) is an essential protein for centriole duplication and cell
division. SAS-6 dimers assemble into a nine-fold ring of diameter 22 nm, from which nine spokes
radiate outward and guide the polymerization of the nine microtubule triplets of the centriole.
Therefore, SAS-6 rings define the characteristic nine-fold symmetry of centrioles, cilia and flagella.
Recently, the assembly of the SAS-6 rings has been reconstituted on flat mica surfaces and monitored
in real time with photothermally-actuated off-resonance tapping high-speed AFM (PORT-HS-AFM)[1]. The assembly curves were analyzed with image processing and a kinetic model for self-assembly(the coagulation-fragmentation equations in the reaction-limited approximation). It was found that thepresence of the surface shifts the reaction equilibrium up by four orders of magnitude compared tothe solution. Using molecular dynamics computer simulations, it was further found that the surface
induces a conformational change in the SAS-6 dimers that allows for ring formation and leads to aresidual asymmetry in the arrangement of the spokes. The PORT-HS-AFM experiments also revealed
that rings can form with other than the nine-fold symmetry. Using Brownian dynamics computer
simulations, it was found that the selectivity for the nine-fold symmetry competes with reversibility of
the assembly pathway, which is critical to avoid kinetic trapping [2]. Together, these studies
demonstrated the intricate interplay between surfaces and protein assembly, and how it can bequantitatively analyzed by combining high-speed AFM with computer simulations.

Honors2021 – 2022
2020 – present
2019 – present
2017
2013
2011 – 2013

Fellow of the Marsilius Kolleg Heidelberg
Member of the biophysics panel of the German Research Foundation

References1.

Fellow of the Max Planck School Matter to Life
Outstanding teaching award of the Department of Physics and Astronomy
Outstanding Referee of the American Physical Society

2.

Banterle, Niccolò, et al. "Kinetic and structural roles for the surface in guiding SAS-6 self-assembly to direct centriole architecture." Nature
Communications 12.1 (2021): 6180.
Gomez Melo, Santiago, et al. "Grand canonical Brownian dynamics simulations of adsorption and self-assembly of SAS-6 rings on asurface." The Journal of Chemical Physics 158.8 (2023).

Speaker of the division Biological Physics of the German Physical Society

Publications
1.
2.
3.
4.
5.

10

Anna Zelená, Johannes Blumberg, Dimitri Probst, Rūta Gerasimaitė, Gražvydas Lukinavičius, Ulrich S. Schwarz, and Sarah Köster. Forcegeneration in human blood platelets by filamentous actomyosin structures. Biophysical Journal, 122(16):3340–3353, 2023.
Oliver M Drozdowski, Falko Ziebert, and Ulrich S Schwarz. Optogenetic control of migration of contractile cells predicted by an active
gel model. Communications Physics, 6(1):158, 2023.
Gonen Golani and Ulrich S Schwarz. High curvature promotes fusion of lipid membranes: Predictions from continuum elastic theory.
Biophysical Journal, 122(10):1868–1882, 2023.
Santiago Gomez Melo, Dennis Wörthmüller, Pierre Gönczy, Niccolo Banterle, and Ulrich Sebastian Schwarz. Grand canonicalBrownian dynamics simulations of adsorption and self-assembly of SAS-6 rings on a surface. The Journal of Chemical Physics, 158(8),2023.
Markus Mund, Aline Tschanz, Yu-Le Wu, Felix Frey, Johanna L Mehl, Marko Kaksonen, Ori Avinoam, Ulrich S Schwarz, and Jonas Ries.
Clathrin coats partially preassemble and subsequently bend during endocytosis. Journal of Cell Biology, 222(3):e202206038, 2023.

11

Clemens M. Franz

https://nanolsi.kanazawa-u.ac.jp/
en/researcher/clemens-franz/clemens-franz-page/

Associate Professor

Investigating Actomyosin Contractility-driven
Adhesion Modulation by ComplementaryHigh-speed AFM and Fluorescence Microscopy

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

Clemens M. Franz

E-mail: clemens.franz@staff.kanazawa-u.ac,jp
Website: https://nanolsi.kanazawa-u.ac.jp/en/researcher/clemens-franz/clemens-franz-page/Research Interests
Atomic Force Microscopy, Fluorescence Microscopy, Cell Biology, Biophysics, Biomaterials

Education
2003

Doctor of Philosophy (PhD) in Cell Biology, University College London, UK

Professional Experience2019 – present
2018 – present
2016 – 20182007 – 20162003 – 2007

Associate Professor, Nano Life Science Institute (WPI-NanoLSI), Kanazawa University, Japan
Assistant Professor, Nano Life Science Institute (WPI-NanoLSI), Kanazawa University, Japan
Senior Lecturer, Institute of Zoology, Karlsruhe Institute of Technology, Germany
Group Leader Nanobiology, Center for Functional Nanostructures, Karlsruhe Institute of Technology, Germany
Postdoctoral Research Fellow, Cellular Machines Group, Max Planck Institute for Molecular Cell Biology and Genetics
and Technical University Dresden, Germany

Honors2014
2000

KIT Research Fellow
Boehringer Ingelheim Fonds Scholar

Publications
1.

12Eroles, M.; Lopez-Alonso, J.; Ortega, A.; Boudier, T.; Gharzeddine, K.; Lafont, F.; Franz, C. M.; Millet, A.; Valotteau, C.; Rico, F.,
Coupled mechanical mapping and interference contrast microscopy reveal viscoelastic and adhesion hallmarks of monocyte
differentiation into macrophages, Nanoscale 2023, 15 [29] 12255-12269

2.

Baumann, H.; Schwingel, M.; Sestu, M.; Burcza, A.; Marg, S.; Ziegler, W.; Taubenberger, A.V.; Muller, D.J.; Bastmeyer, M.; Franz, C.
M., Biphasic reinforcement of nascent adhesions by vinculin, Journal of Molecular Recognition 2023, 6 [36], e3012

3.

Marchesi, A.; Umeda, K.; Komekawa, T.; Matsubara, T.; Flechsig, H.; Ando, T.; Watanabe, S.; Kodera, N.; Franz, C. M., An ultra-wide
scanner for large-area high-speed atomic force microscopy with megapixel resolution, Scientific Reports 2021, 11, 13003

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

Focal adhesions are dynamic integrin adhesion sites where intracellular contractile forces generated
by actin stress fibers are transmitted onto the extracellular environment, thereby driving processes
such as cell migration, tissue invasions, and extracellular matrix (ECM) remodeling, At the same time,
many mechano-sensitive focal adhesion components themselves undergo force-induced
conformational changes and functional regulation. High-speed atomic force microscopy (HS-AFM)
can image such force-induced conformational changes of focal adhesion-associated proteins underphysiological conditions and in real-time. Here, we have applied HS-AFM in combination withfluorescence microscopy to investigate actomyosin contractility-dependent adhesion modulation,
including the tension-driven opening of Ca 2+ channels near mechanically stressed focal adhesion
sites, leading to intracellular Ca 2+ influx, recruitment of Ca2+-binding proteins such as S100A11, andsubsequent focal adhesion disassembly. Furthermore, we have established methods to image
individual integrin receptor-ligand pairs by HS-AFM and show how force-induced conformationalchanges modulate integrin receptor binding strength to the ECM protein laminin. Lastly, by combining
cell deroofing with large-range/high-resolution HS-AFM imaging, we are able to image large
intracellular protein assemblies and even entire organelles down to molecular resolution, while
preserving them in a functional state. In this way, we have generated the first molecularresolution-scale overview images of entire actin stress fibers and analyzed nanostructural and-mechanical changes during myosin II-driven actin stress fiber contraction. Thus, HS-AFM canprovide unique nanoscale structural insight into both intra- and extracellular biomechanical processes
underlying cell/matrix adhesion regulation.

References
1.
2.
3.

Mohammed, T.O.; Lin, Y.R.,; Weissenbruch, K.; Ngo, K.X.; Zhang, Y.; Kodera, N.; Bastmeyer, M.; Miyanari, Y.; Taoka, A.; Franz, C.M.,
S100A11 promotes focal adhesion disassembly via myosin II-driven contractility and Piezo1-mediated Ca2+ entry, bioRxiv 2023
Baumann, H.; Schwingel, M.; Sestu, M.; Burcza, A.; Marg, S.; Ziegler, W.; Taubenberger, A.V.; Muller, D.J.; Bastmeyer, M.; Franz, C. M.,
Biphasic reinforcement of nascent adhesions by vinculin, Journal of Molecular Recognition 2023, 6 [36], e3012
Marchesi, A.; Umeda, K.; Komekawa, T.; Matsubara, T.; Flechsig, H.; Ando, T.; Watanabe, S.; Kodera, N.; Franz, C. M., An ultra-wide
scanner for large-area high-speed atomic force microscopy with megapixel resolution, Scientific Reports 2021, 11, 13003

13

Erik Schäffer

Ultraresolution Picotensiometryof Kinesin Motors

http://www.uni-tuebingen.de/?id=36751

Professor, Doctor
Cellular Nanoscience, ZMBP, University of Tübingen, Germany

Erik Schäffer

E-mail: erik.schaeffer@uni-tuebingen.de
Website: http://www.uni-tuebingen.de/?id=36751

Research Interests
Molecular machines, microtubules and associated proteins, DNA-binding proteins, single-molecule biophysics, optical tweezers

Education
2001

Doctoral degree in physics, University of Konstanz, Germany

Professional Experience
2012 – present
2007 – 20122002 – 2006
1999 – 2001
1999
1998 – 1999
1995 – 1997

Cellular Nanoscience, ZMBP, University of Tübingen, Germany

Simultaneously measuring the nanoscale motion and forces that molecular machines generateprovides insights into how they work mechanically to fulfill their cellular function. To study thesemachines, we developed germanium nanospheres as probes for optical tweezers. With thesehigh–refractive index nanospheres, we have improved the spatiotemporal resolution of opticaltweezers and discovered that the motor kinesin takes 4-nanometer substeps. Further, instead ofdetaching from their microtubule track under load, motors slid back on it, enabling rapid
reengagement in transport. Germanium nanospheres are promising for bioimaging, sensing,
optoelectronics, nanophotonics, and energy storage. For optical trapping, the nanospheres open a
new temporal window by which to uncover hidden dynamics in molecular machines.

Professor of Cellular Nanoscience at the Center for Plant Molecular Biology (ZMBP), University of Tübingen, Germany
Research group leader at the Biotechnology Center (BIOTEC), TU Dresden, Germany
Post-doctoral fellow, Max Planck Institute of Molecular Cell Biology and Genetics (MPI-CBG), Dresden, Germany
Research assistant at the Polymer Chemistry Department, University of Groningen, The Netherlands
Research assistant at the Polymer Science and Engineering Department, University of Massachusetts, Amherst, USA

Reference
1.

S. Sudhakar, M.K. Abdosamadi, T.J. Jachowski, M. Bugiel, A. Jannasch, E. Schäffer, Germanium nanospheres for ultraresolution
picotensiometry of kinesin motors, Science 2021, 371, eabd9944.

Research assistant at the Department of Physics, University of Konstanz, Germany
Research assistant at the Department of Physics, University of Massachusetts, Amherst, USA

Honors
2016
2016
2010
2007
2001

Prize for Bold Science, State of Baden-Württemberg
ERC Proof of Concept, European Research Council (ERC)ERC Starting Grant, European Research Council (ERC)
Emmy-Noether fellowship, German Science Foundation (DFG)DSM Prize for Chemistry and Technology 2001 laureate

Publications

14

1.

Sudhakar, S., Jachowski, T. J., Kittelberger, M., Maqbool, A., Hermsdorf, G. L., Abdosamadi, M. K., and Schäffer, E. Germanium
nanospheres for ultraresolution picotensiometry of kinesin motors. Science 371, eabd9944 (2021)

2.

Ramaiya, A., Roy, B., Bugiel, M., and Schäffer, E. Kinesin rotates unidirectionally and generates torque while walking on microtubules. Proc.
Natl. Acad. Sci. U. S. A. 114, 10894–10899 (2017).

3.

Bormuth, V., Varga, V., Howard, J., and Schäffer, E. Protein friction limits diffusive and directed movements of kinesin motors on microtubules.
Science 325, 870–873 (2009).

15

Masanobu Oshima

Identification of Genotype-linked
Nano-Scale Physical Properties
of Intestinal Tumor Cells

http://genetics.w3.
kanazawa-u.ac.jp/english/

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

Masanobu Oshima

E-mail: oshimam@staff.kanazawa-u.ac.jp
Website: http://genetics.w3.kanazawa-u.ac.jp/english/

Research Interests
Cancer Biology, Mouse Models and Organoid Models for Tumors, Nano-Scale Study for Malignancy

Education
1988

Hokkaido University Graduate School of Veterinary Medicine, Japan

Professional Experience2017 – present
2005 – present
2000 – 2005
1997 – 1999
1992 – 1997
1988 – 1992

Professor, WPI-NanoLSI, Kanazawa University, Japan
Professor, Cancer Research Institute, Kanazawa University, Japan
Associate Professor, Kyoto University Graduate School of Medicne, Japan
Research Associate, Merck Research Laboratories (Merck), USA
Staff Scientist, Banyu Pharmaceutical Co., Ltd., Japan
Staff Scientist, Chugai Pharmaceutical Co., Ltd., Japan

Honors2015
2012

Accumulation of driver gene mutations is responsible for colon cancer development. However, it has
not yet been fully understood, which driver mutations induce each process of malignant progression,
such as invasion, dissemination, and metastasis. To examine the malignant progression mechanism,
we generated mouse models that carry driver mutations in driver genes, Apc (A), Kras (K), Tgfbr2 (T),Trp53 (P), and Fbxw7 (F) in various combinations, and established tumor-derived organoids fromthese models. Comprehensive histological analyses of in vivo tumors clarified the link between
genotype and phenotypes. Namely, AT and AP double mutations induce submucosal invasion, AKT
and AKP triple mutations causes advanced malignancy like EMT and intravasation, and AKTP and
AKTPF multiple combinations drive liver metastasis at high frequency. Interestingly, we found that
non-metastatic organoids can metastasize when co-migrated with metastatic cells by “polyclonalmechanism” . We next examined physical properties of the membrane surface of theses organoid
cells by using high-speed ion conductance microscope (HS-SICM) time-lapse imaging. Notably,highly metastatic AKTP and AKTPF cells showed specific micro-ridges on the cell surface, and thesestructures were actively and continuously moving. Moreover, the stiffness of the metastatic AKTP and
AKTPF cells was significantly lower compared with non-metastatic and benign tumor cells. Such
results from organoid studies will be important for understanding the mechanism of malignant
progression of cancer.

The Commendation for Science and Technology by the Ministry of Education, Culture, Sports, Science and Technology (MEXT)

References

JCA-Mauvernay Award, Japanese Cancer Association (JCA)

1.

Publications

16

Nano Life Science Institute (WPI-NanoLSI), Kanazawa University, Japan
Cancer Research Institute, Kanazawa University, Japan

1.

Wang D, Sun L, Okuda S, Yamamoto D, Nakayama M, Oshima H, Saito H, Kouyama Y, Mimori K, Ando T, Watanabe S, and Oshima
M. Nano-scale physical properties characteristic to metastatic intestinal cancer cells identified by high-speed scanning ion
conductance microscope. Biomaterials, 280: 121256, 2022.

2.

Kok SY, Oshima H, Takahashi K, Nakayama M, Murakami K, Ueda HR, Miyazono K, and Oshima M. Malignant subclone drives metastasisof genetically and phenotypically heterogenous cell clusters through fibrotic niche generation. Nat Commun, 12 (1): 863, 2021.

3.

Takeda H, Kataoka S, Nakayama M, Ali MAE, Oshima H, Yamamoto D, Park JW, Takegami Y, An T, Jenkins NA, Copeland NG, andOshima M. CRISPR-Cas9 mediated gene knockout in intestinal tumor organoids provides functional validation for colorectal cancer drivergenes. Proc Natl Acad Sci USA, 116: 15635-15644, 2019.

4.

Oshima H, Kok SY, Nakayama M, Murakami K, Voon DC, Kimura T, and Oshima M. Stat3 is indispensable for damage-induced cryptregeneration but not for Wnt-driven intestinal tumorigenesis. FASEB J, 33: 1873-1886, 2019.

2.
3.

4.

Sakai E, Nakayama M, Oshima H, Kouyama Y, Niida A, Fujii S, Ochiai A, Nakayama KI, Mimori K, Suzuki Y, Hong CP, Ock CY, Kim SJ,
and Oshima M. Combined mutation of Apc, Kras and Tgfbr2 effectively drives metastasis of intestinal cancer. Cancer Res. 2018, 78 (5),1334-1346,.
Kok SY, Oshima H, Takahashi K, Nakayama M, Murakami K, Ueda HR, Miyazono K, and Oshima M. Malignant subclone drives
metastasis of genetically and phenotypically heterogenous cell clusters through fibrotic niche generation. Nat. Commun. 2021, 12 (1), 863.
Wang D, Sun L, Okuda S, Yamamoto D, Nakayama M, Oshima H, Saito H, Kouyama Y, Mimori K, Ando T, Watanabe S, and Oshima M.
Nano-scale physical properties characteristic to metastatic intestinal cancer cells identified by high-speed scanning ion conductancemicroscope. Biomaterials 2022, 280, 121256.
Wang D, Nguyen HG, Nakayama M, Oshima H, Sun L, Oshima M, and Watanabe S. Mapping nanomechanical properties of basal
surfaces in metastatic intestinal 3D living organoids with high-speed scanning ion conductance microscopy. Small 2023, 19 (9), e2206213.

17

Anna Taubenberger

https://tu-dresden.de/cmcb/biotec/
forschungsgruppen/taubenberger

Quantifying Tumor Spheroid Mechanical Properties
in Confining 3D Microenvironments

Group leader Lab of Oncomechanics
Center for Molecular and Cellular Bioengineering (CMCB), TU Dresden, Germany

Anna Verena Taubenberger

E-mail: Anna.taubenberger@tu-dresden.de
Website: https://tu-dresden.de/cmcb/biotec/forschungsgruppen/taubenberger

Research Interests
Cell and tissue mechanics, tumor microenvironment, bioengineered in vitro models

Education
2009
2005

Doctor of Engineering, Technische Universität Dresden, Germany
Diploma (Dipl.-Ing.) Engineering, Technische Universität Dresden, Germany

Professional Experience2020 – present2013 – 20202009 – 2012

Group leader, MSNZ fellow, CMCB TU Dresden
PostDoctoral Research fellow, CMCB TU Dresden
PostDoctoral Research fellow, Queensland University of Technology, Brisbane, Australia

Honors
2020
2019
2011
2010

Mildred Scheel Nachwuchszentrum (MSNZ) Medical Scientist

Biotechnology Center of the Technische Universität Dresden, Germany

Tumor growth and invasion occurs in a mechanically altered tumor microenvironment, which isthought to contribute to cancer progression. Effects of matrix stiffness on tumor cells are well studied ina systematic manner using bioengineered 3D tumor in-vitro models. Previous studies indicatethat tumor spheroids change their growth and invasion behavior when grown in stiffened
microenvironments. Still, it remains poorly understood how the cells mechanically adapt to confining
3D microenvironments. Here we cultured single breast and pancreatic cancer cells to form tumor
spheroids within ECM-mimicking biohybrid hydrogels of defined mechanical properties. Systematic
modulation of hydrogel stiffness and/or degradability were associated with changes in cellmorphology, gene expression and spheroid growth. Brillouin microscopy and atomic force
microscopy revealed that spheroids altered their mechanical properties under confinement at thecellular and multicellular scale and when adopting an invasive phenotype. Drugs interfering withcell-cell junctions and intermediate filaments, but not F-actin filaments or microtubules affected themechanical phenotype of tumor spheroids measured in situ. Taken together, our study provides
insights into how tumor cells adapt their mechanical properties to microenvironment stiffness andconfinement and when forming multicellular and invasive structures, which is relevant to tumor
formation and progression.

References

Poster Prize EACR
Early Career Research Fellowship QUT

1.

DFG PostDoc fellowship2.

Mahajan V, Beck T, Gregorczyk P, Ruland A, Alberti S, Guck J, Werner C, Schlüßler R, Taubenberger AV. Mapping tumor spheroid
mechanics in dependence of 3D microenvironment stiffness and degradability by Brillouin microscopy. Cancers 13:5549 (2021).
Taubenberger A, Girardo S, Träber N, Fischer Friedrich E, Kräter M, Wagner K, Kurth T, Richter I, Haller B, Binner M, Hahn D, Freudenberg
U, Werner C, Guck J, 3D microenvironment stiffness regulates tumor spheroid growth and mechanics via p21 and ROCK. Advanced
Biosystems 391:1900128–16 (2019).

Publications

18

1.

Abuhattum S, Kotzbeck P, Schlüßler R, Harger A, Ariza Schellenberger A, Kim K, Escolano JC, Müller T, Braun J, Wabitsch M, Tschöp
M, Sack I, Brankatschk M, Guck J, Stemmer K, Taubenberger AV. Adipose cells and tissues soften with lipid accumulation while indiabetes adipose tissue stiffens. Sci Rep 12: 10325 (2022).

2.

Mahajan V, Beck T, Gregorczyk P, Ruland A, Alberti S, Guck J, Werner C, Schlüßler R, Taubenberger AV. Mapping tumor spheroid
mechanics in dependence of 3D microenvironment stiffness and degradability by Brillouin microscopy. Cancers 13:5549 (2021).

3.

Jäschke A, Jacobi A, Lawrence MG, Risbridger GP, Frydenberg M, Williams ED, Vela I, Hutmacher DW, Bray LJ, Taubenberger A.
Cancer-associated fibroblasts of the prostate promote a compliant and more invasive phenotype in benign prostate epithelial cells.
Mater Today Bio 100073 (2020).

19

Satoru Okuda

Versatile Computational Modeling of
3D Multicellular Dynamics and Application
to Collective Cell Migration

http://mechgen.jp

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

Satoru Okuda

E-mail: satokuda@staff.kanazawa-u.ac.jp
Website: http://mechgen.jp

Research Interests
Biomechanics, Multicellular dynamics, Morphogenesis, Computational mechanics

Education
2013

Doctor of Engineering, Department of Micro Engineering, Kyoto University, Japan

Professional Experience2019 – present
2016 – 2021
2016 – 20182013 – 2016

Associate Professor, Nano Life Science Institute, Kanazawa University, Japan
PRESTO Researcher, Japan Science and Technology Agency, Japan

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

In embryogenesis and cancer invasion, cells migrate collectively as a cluster in 3D tissues. Many
studies have elucidated the mechanisms of either individual or collective cell migration on 2D
substrates; however, it remains unclear how cells migrate collectively as a cluster in 3D tissues. To
address such 3D multicellular dynamics, we developed a versatile 3D vertex model and applied it tocollective cell migration in 3D space. Numerical simulations revealed that polarized interfacial
tension enables cells to migrate collectively as a cluster through a 3D tissue. In this mechanism, theinterfacial tension induces unidirectional flow of each cell surface from the front to the rear along thecluster surface. Importantly, multiple migratory modes were induced depending on the strength ofpolarity, adhesion, and noise, i.e., cells migrated either as single cells, as a cluster, or aligned likebeads on a string, as occurs in embryogenesis and cancer invasion. These results suggest that thesimple expansion and contraction of cell-cell boundaries enables cells to move directionally andgenerate the variety of collective migrations observed in living systems.

Special Postdoctoral Researcher, RIKEN Center for Developmental Biology, Japan
Researcher, RIKEN Center for Developmental Biology, Japan

References

Honors
2021
2020
2019
2017
2017

Seguchi Award, Japan Society of Mechanical Engineers

1.
2.

Rising Star Award, Japan Science and Technology Agency PRESTO Area
Japan Society of Mechanical Engineers Award (Paper)
Yamaguchi Medal, Asian-Pacific Association for Biomechanics
Incentive Award (Research), Japan Society of Mechanical Engineers

3.
4.

S Okuda*, K Sato*, "Polarized interfacial tension induces collective migration of cells, as a cluster, in a 3D tissue," Biophysical Journal 121(10): 1856-1867 (2022)
S Okuda*, K Fujimoto, "A Mechanical Instability in Planar Epithelial Monolayers Leads to Cell Extrusion," Biophysical Journal 118(10):2549-2560 (2020)
S Okuda*, E Kuranaga, K Sato*, "Apical junctional fluctuations lead to cell flow while maintaining epithelial integrity," Biophysical Journal
116 (6): 1159-1170 (2019)
S Okuda*, N Takata, Y Hasegawa, M Kawada, Y Inoue, T Adachi, Y Sasai, M Eiraku*, "Strain-triggered mechanical feedback in
self-organizing optic cup morphogenesis," Science Advances 4(11): eaau1354 (2018)

Publications

20

1.

S Okuda*, T Hiraiwa, "Modelling contractile ring formation and division to daughter cells for simulating proliferative multicellular
dynamics," The European Physical Journal E 46: 56 (2023)

2.

R Matsuzawa, A Matsuo, S Fukamachi, S Shimada, M Takeuchi, T Nishina, P Kollmannsberger, R Sudo, S Okuda*, T Yamashita*,"Multicellular dynamics on structured surfaces: Stress concentration is a key to controlling complex microtissue morphology on
engineered scaffolds," Acta Biomaterialia 166: 301-316 (2023)

3.

S Okuda*, T Hiraiwa, "Long-term adherent cell dynamics emerging from energetic and frictional interactions at the interface," PhysicalReview E 107 (3): 034406 (2023)

21

Martin Falcke

On Multistability and Constitutive Relationsof Cell Motion on Fibronectin lanes

https://www.mdc-berlin.de/

Martin Falcke

Professor, Doctor
Max Delbrück Center for Molecular Medicine Berlin, Germany

Max Delbrück Center for Molecular Medicine Berlin

E-mail: Martin.falcke@mdc-berlin.de
Website: https://www.mdc-berlin.de/Research Interests
Biophysics, Statistical Physics, Non-linear Dynamics

Education
2005
1995
1987

Habilitation: Theoretical Physics, Freie Universität Berlin
Promotion Dr. rer. nat.: Theoretical Physics, Technische Universität Berlin
Diploma Theoretical Physics, Technische Universität Chemnitz

Professional Experience
2014
2008 – present
2001 – 2008
1997 – 98 and 2000
1995 – 97, 99
1995
1990 – 1995
1987 – 1990

apl. Professor, Dept. of Physics, Humboldt University Berlin
Research group leader, Max Delbrück Center for Molecular Medicine Berlin
Research assistant, Hahn-Meitner-Institute Berlin
Research assistant, Max-Planck-Institute for the Physics of Complex Systems Dresden
Post doc, Postdoc positions at the Institute for Nonlinear Science and the Physics Department of the University of California San Diego, USA
Post doc, Chemical Engineering Department of the University of Virginia in Charlottesville, USA
Research assistant, TU Berlin
Engineer for control systems, VEB Chemieanlagenbau Leipzig-Grimma, Direktionsbereich Generallieferant, Karl-Marx-Stadt

References

Honors2003

Cell motility on flat substrates exhibits coexisting steady and oscillatory morphodynamics, the biphasic
adhesion-velocity relation, and the universal correlation between speed and persistence (UCSP) as
simultaneous observations common to many cell types. Their universality and concurrency suggest aunifying mechanism causing all three of them. Stick-slip models for cells on 1dimensional lanes
suggest multistability to arise from the non-linear friction of retrograde flow. This study suggests a
mechanical mechanism controlled by integrin signalling on the basis of a biophysical model andanalysis of trajectories of MDA-MB-231 cells on Fibronectin lanes which additionally explains theconstitutive relations(1, 2). The experiments exhibit cells with steady or oscillatory morphodynamics
and either spread or moving with spontaneous transitions between the dynamic regimes, spread andmoving and spontaneous direction reversals. Our biophysical model is based on the force balance atthe protrusion edge, the noisy clutch of retrograde flow and a response function of friction andmembrane drag to integrin signaling. The theory reproduces the experimentally observed cell states,
characteristics of oscillations and state probabilities. Analysis of experiments with the biophysicalmodel establishes a stick-slip oscillation mechanism, explains multistability of cell states and thestatistics of state transitions. It suggests protrusion competition to cause direction reversal events, thestatistics of which explain the UCSP. The effect of integrin signalling on drag and friction explains theadhesion-velocity relation and cell behavior at Fibronectin density steps. The dynamics of ourmechanism are non-linear flow mechanics driven by F-actin polymerization and shaped by the noisy
clutch of retrograde flow friction, protrusion competition via membrane tension and drag forces.
Integrin signalling controls the parameters of the mechanical system.

1.
Schrödinger Prize for interdisciplinary research of the Donors' Association for the Promotion of
the Sciences and Humanities in Germany 2003, together with J.D. Lechleiter and P. Camacho

2.

Amiri B, Heyn JCJ, Schreiber C, Rädler JO, & Falcke M (2023) On multistability and constitutive relations of cell motion on fibronectin lanes.
Biophysical Journal 122(5):753-766.
Schreiber C, Amiri B, Heyn JCJ, Rädler JO, & Falcke M (2021) On the adhesion–velocity relation and length adaptation of motile cells on
stepped fibronectin lanes. Proceedings of the National Academy of Sciences 118(4):e2009959118.

Publications

22

1.

Amiri B, Heyn JCJ, Schreiber C, Rädler JO, & Falcke M (2023) On multistability and constitutive relations of cell motion on fibronectinlanes. Biophysical Journal 122(5):753-766.

2.

Schreiber C, Amiri B, Heyn JCJ, Rädler JO, & Falcke M (2021) On the adhesion–velocity relation and length adaptation of motile cells
on stepped fibronectin lanes. Proceedings of the National Academy of Sciences 118(4):e2009959118.

3.

Dimchev G, et al. (2020) Lamellipodin tunes cell migration by stabilizing protrusions and promoting adhesion formation. Journal of Cell
Science 133(7):jcs239020.

4.

Dolati S, et al. (2018) On the relation between filament density, force generation, and protrusion rate in mesenchymal cell motility.
Molecular Biology of the Cell 29(22):2674-2686.
23Mark J. MacLachlan

Supramolecular Chemistry Using Bionanomaterials
https://www.chem.ubc.ca/
mark-maclachlan

Mark J. MacLachlan

Professor
University of British Columbia, Vancouver, Canada
Nano Life Science Institute (WPI-NanoLSI), Kanazawa University, Japan

E-mail: mmaclach@chem.ubc.ca
Website: https://www.chem.ubc.ca/mark-maclachlan

Research Interests
supramolecular chemistry, materials chemistry, biomaterials, photonic materials

Education
1999
1995

PhD, Department of Chemistry, University of Toronto, Canada
BSc, Department of Chemistry, University of British Columbia, Canada

Professional Experience2017 – present
2016 – 2022
2014 – 2022
2011 – present
2007 – 2011
2001 – 2007
1999 – 2001

University of British Columbia, Vancouver, Canada
Nano Life Science Institute (WPI-NanoLSI), Kanazawa University, Japan

Bionanomaterials, such as cellulose nanocrystals (CNCs), are attractive for incorporation into newmaterials as they are extracted from renewable resources, they are biodegradable, and theirproperties can be changed through facile surface modification. Furthermore, CNCs are known toorganize into a chiral nematic liquid crystalline phase in water. Upon drying, their order can beretained in solid films, with the organization resembling a helical stacking of the nanocrystals. Thesefilms with chiral nematic order display photonic properties (they appear iridescent) when the helicalpitch of the structure is on the order of the wavelength of visible light. By deliberately changing thehelical pitch of the structure, one can readily tune the wavelength of light reflected by the films.
In this presentation, I will discuss our group’s efforts to construct new photonic materials using CNCs
as an active component or a template. In particular, we have been constructing elastomers andshape-memory composite materials whose photonic properties change upon mechanical stimulation.

Professor, Nano Life Science Institute (WPI-NanoLSI), Kanazawa University
Associate Dean for Research & Graduate Studies, Faculty of Science, UBC
Director, NSERC CREATE Training Program in Nanomaterials (NanoMat)
Professor, UBC
Associate Professor, UBC
Assistant Professor, UBC
NSERC Postdoctoral Fellow, MIT, Cambridge, USA

References
1.
2.
3.
4.

Honors2015 – 2029
2016
2016
2014
2014
2013
2013
2012-2014

5.

Boott, C. E.; Shi, L.; Li, Z.; Hamad, W. Y.; MacLachlan, M. J. “A Facile Photoinitiated Polymerization Route for the Preparation of Photonic
Elastomers with Chiral Nematic Order” Liq. Cryst. 2023, in press.
Xu, M; Xu, Z.; Soto, M. A.; Xu, Y.-T.; Hamad, W. Y.; MacLachlan, M. J. “Mechanically Responsive Circularly Polarized Luminescence from
Cellulose Nanocrystal-based Shape-Memory Polymers” Adv. Mater. 2023, in press.
Yurtsever, A.; Wang, P.-X.; Priante, F.; Jaques, Y. M.; Miyazawa, K.; MacLachlan, M. J.; Foster, A. S.; Fukuma, T. “Molecular insights at the
crystalline cellulose-water interfaces by three-dimensional atomic force microscopy” Sci. Adv. 2022, 8, eabq0160.
Boott, C. E.; Soto, M. A.; Hamad, W. Y.; MacLachlan, M. J. “Shape-Memory Photonic Thermoplastics from Cellulose Nanocrystals” Adv.
Funct. Mater. 2021, 31, 2103268.
Boott, C. E.; Tran, A.; Hamad, W. Y.; MacLachlan, M. J. “Cellulose Nanocrystal Elastomers with Reversible Visible Color” Angew. Chem. Int.
Ed. 2020, 59, 226-231.

Tier 1 Canada Research Chair in Supramolecular Materials
Fellow of the Royal Society of Chemistry (UK)Award for Excellence in Materials Chemistry, Canadian Society for Chemistry
Steacie Prize
Fellow of the Royal Society of Canada
Rutherford Memorial Medal in Chemistry, Royal Society of Canada
JSPS Invitational Fellowship for Research
E.W.R. Steacie Memorial Fellowship

Publications
1.
2.
3.
24

Cho, K.; Andrew, L. J.; MacLachlan, M. J. “Uniform Growth of Nanocrystalline ZIF-8 on Cellulose Nanocrystals: Useful Template forMicroporous Organic Polymers” Angew. Chem. Int. Ed. 2023, 62, e202300960.
Soto, M. A.; Carta, V.; Suzana, I.; Patrick, B. O.; Lelj, F.; MacLachlan, M. J. “Cycling a Tether into Multiple Rings: Pt-Bridged
Macrocycles for Differentiated Guest Recognition, Pseudorotaxane Transformations, and Guest Capture and Release” Angew. Chem.
Int. Ed. 2023, 62, e202216029.
Xu, Y.-T.; Li, J.; MacLachlan, M. J. “Stable Graphene Oxide Hydrophobic Photonic Liquids” Nanoscale Horizons 2022, 7, 185-191.

25

Martin Bastmeyer

3D Cellular Microenvironments
to Study Cell Mechanics

https://znbio.zoo.kit.edu/

Professor, Doctor
Zoological Institute, Cell- and Neurobiology,
Karlsruhe Institute of Technology (KIT), Germany

Martin Bastmeyer

E-mail: bastmeyer@kit.edu
Website: https://znbio.zoo.kit.edu/

Research Interests
Cell mechanics, Stem Cell Biology, Advanced Microscopy, Developmental Neurobiology

Education
1989
1984

PhD in Biology, MPI Tübingen and University Kaiserslautern
Diploma in Biology, University Kaiserslautern

Professional Experience
2021 – present
2004 – present
2001 – 2004
1998 – 2001
1993 – 1994
1988 – 1998

Director Institute of Biological and Chemical Systems - Biological Information Processing (IBCS-BIP) at KITProfessor for Cell- and Neurobiology, Karlsruhe Institute of Technology (KIT), Germany
Professor for Neurobiology, University Jena, Germany
Independent Research Group Leader, University Konstanz, Germany
Postdoc, Salk Institute, San Diego, USA
Postdoc, MPI Tübingen and University Konstanz, Germany

Zoological Institute, Cell- and Neurobiology, Karlsruhe Institute of Technology (KIT), Germany

Cell behavior and differentiation are not only influenced by biochemical cues but also by physicalproperties like adhesive geometry, topography, and stiffness of the 3D extracellular environment.
Correspondingly, cells exert, sense, and respond to physical forces by a variety of mechanisms. In thistalk I will discuss how 3D laser nanoprinting can be applied to design 3D cellular microenvironmentsin the µm range with defined geometries and adjustable flexibility [1,2]. By varying the adhesion
geometry, we study the interplay between extracellular physical factors and cell function in asystematic approach. Since the elastic modulus of the scaffold material varies between E=140-350
MPa, measurements of cell adhesion forces (typically in the range of 50-100 nN) in relation toadhesion geometry are also feasible. Furthermore, we are developing stimuli-responsive resists for 3D
laser nanoprinting [3]. These hydrogels change their mechanical properties upon an external stimulus
and thus have great potential for applications to study cellular mechanobiology. We have
manufactured a miniaturized device that can be used to mechanically stretch single cells in awell-defined temporal and spatial manner and to simultaneously measure cellular contraction forces
during the experiment [4] (Fig. 1). In combination with CRISPR/Cas9-generated knockout cell linesof proteins involved in cellular force production (e.g., non-muscle myosin II isoforms, a-actinin
isoforms) this system will allow to study the molecular basis and regulation of force generation by
non-muscle cells [5].

Honors
2018
2016
2014
1998

1

BRIDGE Fellowship of the Japanese Society for the Promotion of Sciences (JSPS)

2

3

4

2

5

3

4

Erwin-Schrödinger-Prize for Interdisciplinary Research
Advanced Researcher Fellowship Japanese Society for the Promotion of Sciences (JSPS)

1

Heisenberg Fellowship of the DFG
InitialStretchResponse

Release5

Response

Publications

26

1.

K Weißenbruch, J Grewe, M Hippler, M Fladung, M Tremmel, K Stricker, US Schwarz, M Bastmeyer (2021) Distinct roles of
nonmuscle myosin II isoforms for establishing tension and elasticity during cell morphodynamics. eLife 10: doi: 10.7554/eLife.71888.

References2.

M Hippler, K Weißenbruch, K Richler, ED Lemma, M Nakahata, B Richter, C Barner-Kowollik, Y Takashima, A Harada, E Blasco, M
Wegener, M Tanaka, M Bastmeyer (2020) Mechanical stimulation of single cells by reversible host-guest interactions in 3D
microscaffolds. Sci Adv. 6(39):eabc2648. doi: 10.1126/sciadv.abc2648.

1.

3.

M Hippler, E Blasco, J. Qu, M Tanaka, C Barner-Kowollik, M Wegener, M Bastmeyer (2019) Controlling the shape of 3D
microstructures by temperature and light. Nat Commun. 10: 232. doi: 10.1038/s41467-018-08175-w.

4.

B Richter, V Hahn, S Berthels, TK Claus, M Wegener, G Delaittre, C Barner-Kowollik and M Bastmeyer (2017) Guiding cell attachment
in three-dimensional microscaffolds selectively functionalized with two distinct adhesion proteins. Adv. Mater. 29: DOI:
10.1002/adma.201604342

5.

F Klein, T Striebel, J Fischer, Z Jiang, CM Franz, G von Freymann, M Wegener and M Bastmeyer (2010) Elastic fully three-dimensional
microstructure scaffolds for cell force measurements. Adv. Mater. 22: 868-871.

2.
3.
4.
5.

K Weißenbruch et al. (2022) Micro-scaffolds as synthetic cell niches: recent advances and challenges. Curr Opin Biotechnol 73: 290-299.
doi: 10.1016/j.
C Barner-Kowollik et al. (2017) 3D Laser Micro- and Nano-Printing: Challenges for Chemistry. Angew. Chem. Int. Ed. 56: 15828-15845,
doi: 10.1002/anie.201704695
CA Spiegel et al. (2020) 4D Printing at the Microscale. Adv Func Mater 30: doi: 10.1002/adfm.201907615
M Hippler et al. (2020) Mechanical stimulation of single cells by reversible host-guest interactions in 3D microscaffolds. Sci Adv. 6(39):
eabc2648. doi: 10.1126/sciadv.abc2648.
K Weißenbruch et al. (2021) Distinct roles of nonmuscle myosin II isoforms for establishing tension and elasticity during cell morphodynamics.
eLife 10: doi: 10.7554/eLife.71888.

27

Svetlana Santer

From Molecular Actuation
to Microscopic Motion

http://exph.physik.uni-potsdam.de/Professor, Doctor
Department of Smart Soft Matter,
Institute of Physics and Astronomy, University of Potsdam, Germany

Svetlana Santer

E-mail: santer@uni-potsdam.de
Website: http://exph.physik.uni-potsdam.de/Research Interests
Light responsive polymers and nano-objects, physics and chemistry at interfaces, functional thin polymer films, manipulation of nano-objects

Education
1997-2000
1989-1996

PhD (Dr. rer.nat.), at the University of Ulm, Germany
Study of physics at the St.-Petersburg State University, Russia

Professional Experience2009 – present
2008 – 20092007
2003 – 2007
2001 – 2003
2000 – 2001

Professor for Experimental Physics, University of Potsdam, Germany
Junior Fellow at the Freiburg Institute of Advanced Studies, Freiburg, Germany
Habilitation in Microsystem Technology (MST) at IMTEK, Freiburg, Germany
Research Assistant at the Institute for Microsystem Technology (IMTEK), Freiburg, Germany
Group Leader “Interface Analysis” at the Freiburger Materialforschungszentrum (FMF), Germany
PostDoc at IMTEK, Freiburg, Germany

Honors
2007
2004
2003
1995

Heisenberg stipend, DFG
Eliteförderprogramm für Postdoktorandinnen und Postdoktoranden, Landesstiftung Baden-Württemberg

Department of Soft Smart Matter, Institute of Physics and Astronomy, University of Potsdam, Germany

Azobenzene molecules can be considered as molecular actuators that convert optical energy inmechanical work. In this talk I will show several interesting examples where azobenzene can be usedin order to actuate matter on large time and length scales. In all these examples the azobenzene
plays the role of a transducer that mediates between different states of size, shape, position andinterfacial energy of several nano-scale soft materials.
In the first part of my talk I will show how using a home made set-up combining an atomic force
microscope (AFM) and two-beam interferometry. It is possible to address two major points
concerning the experimental efforts in understanding surface relief grating (SRG) formation inazobenzene containing polymers: (i) how is the orientation of the electric field vector within theinterfering electromagnetic fields related to the topographical pattern within the SRG; (ii) how can
one measure locally the opto-mechanical forces emerging during topography change. We will
discuss three distinct systems: polymer films, polymer brushes, and azobenzene containing polymer
nanoparticles. [1-4]
In the second part of my talk I will show how, using azobenzene containing surfactant, [5] one canmanipulate microparticles and even induce their self-propulsion when trapped at a solid/liquidinterface. The physical origin of this genuine behavior is related to the phenomenon of light driven
diffusioosmosis (LDDO). [6,7] We will discuss how to establish light-driven hydrodynamics as a usefuland versatile tool for investigating collective motion of self-propelled particles and aggregation. At
the very end of my talk I will present further examples of azo-induced actuation of soft matter.
Microgel particles made light sensitive with azo-surfactants can change their volume (growing or
shrinking) by up to a factor of 8 and LCST point between 32°C and 85°C in response to illumination
with two different wavelengths. [8, 9]Margarethe von Wrangell Habilitationsstipendium
Soros Preis for young researchers for the work on “Interaction of Molecule DNA with Antitumor Compounds

Publications
1.

Polarization controlled fine structure of diffraction spots from an optically induced grating, J. Jelken, C. Henkel, S. Santer*, Appl. Phys.
Lett. 2020, 116, 051601.

2.

Light induced reversible structuring of photosensitive polymer films, J. Jelken, S. Santer* RSC Adv. 2019, 9, 20295.

3.

Remote control of soft nano-objects by light using azobenzene containing surfactants, S. Santer, J. Phys. D Appl. Phys. 2017, 51,
013002.

References
1.
2.
3.
4.
5.
6.
7.
8.
9.

28

Jelken, J.; Santer, S. RSC Advances, 9 (2019) 20295.
Di Florio, G.; Brundermann, E.; Yadavalli, N.S.; Santer, S.A.; Havenith, M. Nano Letters, 14 (2014) 5754.
Loebner, S.; Lomadze, N.; Kopyshev, A.; Koch, M.; Guskova, O.; Grenzer Saphiannikova, M.; Santer, S. A. J. Phys. Chem. B, 122 (2018)2001.
Lomadze, N.; Kopyshev, A.; Bargheer, M.; Wollgarten, M.; Santer, S. Scientific Reports, 7 (2017) 8506.
Santer, S. J. Phys. D: Applied Physics, 51 (2017) 013002.
Feldmann, D.; Maduar S.R.; Santer, M.; Lomadze, N.; Vinogradova O.I.; Santer, S. Scientific Reports, 6 (2016) 36443.
Arya, P.; Umlandt, M.; Jelken, J.; Feldmann, D.; Lomadze, N.; Asmolov , E. S.; Vinogradova, O. I.; Santer, S. A. The European Physical Journal
E, 44(50) (2021), 1-10.
Schimka, S.; Gordievskaya, Y.D.; Lomadze, N.; Lehmann, M.; von Klitzing, R.; Rumyantsev, A.M.; Kramarenko, E.; Santer, S. J. Chem. Phys.,
147 (2017) 031101.
Jelken J.; Jung S.; Lomadze N.; Gordievskaya Y. D.; Kramarenko E. Y.; Pich A.; Santer S. Adv. Funct. Mater., (2021), 2107946.

29

Shinji Watanabe

https://researchmap.jp/swsw

Associate Professor, Doctor

Time-Resolved Imaging of Nanomechanics andNanostructures of Living Cells with
Scanning Ion Conductance Microscopy

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

Shinji Watanabe

E-mail: wshinji@se.kanazawa-u.ac.jp
Website: https://researchmap.jp/swsw

Research Interests
Scanning probe microscopy, Nanoscience, Nanoscale measurement technologies.

Education
2006

Doctor of Science, Graduate School of Science and Technology, Niigata University, Japan

Professional Experience2020 – present2017 – 20202012 – 20172009 – 20122007 – 20092007
2006 – 2007
2006

Associate Professor, Nano Life Science Institute (WPI-NanoLSI), Kanazawa University, Japan
Assistant Professor, Nano Life Science Institute (WPI-NanoLSI), Kanazawa University, Japan
Assistant Professor, Bio-AFM Frontier Research Center, Kanazawa University, Japan
JST-ERATO Postdoctoral Fellow, Tohoku University, Japan
Assistant Professor, Center for the Advancement of Higher Education, Tohoku University, Japan
Assistant Professor, Department of Physics, Tohoku University, Japan

30

Scanning ion conductance microscopy (SICM) is a kind of scanning probe microscopy technique,which uses a glass nanopipette as a probe to detect a tiny ion current through the nanopipette tip
opening for nanoscale imaging. SICM has been utilized to observe nanostructures of living cellsurfaces and their dynamics with low invasiveness in measurements. Recently, SICM has been
advanced to examine the nanomechanical properties of living cell surfaces, allowing us to provide
elasticity maps with nanometer-scale resolution. However, there is still a need to improve image rateand spatial resolution for diverse applications of nanobiosciences. In addition, improvement inaccessing complicated structures, such as fragile tissues, has also been desired in SICM. We haverecently developed a high-speed-scan type of SICM (HS-SICM). With the developed technique, wehave simultaneously visualized nanostructures and elasticity distributions of living cancer cells withdifferent malignancies. In addition, dynamic information of SICM images allows us to suggest
quantitative differences in cellular states as the difference in physical parameters, such as roughness,
mobility, and elasticity. We also applied HS-SICM to investigate nanostructures andnanomechanical properties of three-dimensional living organoids. By developing a simple method toembed organoids in collagen gels, we successfully visualized multiple nano and microstructures andelasticity distributions and their dynamics.

JST- SORST Postdoctoral Fellow, Niigata University, Japan
JST-CREST Postdoctoral Fellow, Niigata University, Japan

Publications
1.

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

Wang, D.; Gia Han., N.; Nakayama, M.; Oshima, H.; Sun, L.; Oshima M.; Watanabe, S.; Mapping Nanomechanical Properties ofBasal Surfaces in Metastatic Intestinal 3D Living Organoids with High-Speed Scanning Ion Conductance Microscopy, Small, 2022,
2206213.

2.

Wang, D.; Sun, L.; Okuda, S.; Yamamoto, D.; Nakayama, M.; Oshima. H; Saito, H.; Kouyama, Y.; Mimori, K.; Ando, T.; Watanabe, S.;
Oshima, M., Nano-scale physical properties characteristic to metastatic intestinal cancer cells identified by high-speed scanning ion
conductance microscope. Biomaterials 2022, 280, 121256

3.

Watanabe, S.; Kitazawa, S.; Sun, L.; Kodera, N.; Ando, T.; Development of High-Speed Ion Conductance Microscopy, Rev. Sci.
Instrum. 2019, 90(12), 123704.

References
1.
2.

3.

Wang, D.; Gia Han., N.; Nakayama, M.; Oshima, H.; Sun, L.; Oshima M.; Watanabe, S.; Mapping Nanomechanical Properties of Basal
Surfaces in Metastatic Intestinal 3D Living Organoids with High-Speed Scanning Ion Conductance Microscopy, Small, 2022, 2206213.
Wang, D.; Sun, L.; Okuda, S.; Yamamoto, D.; Nakayama, M.; Oshima. H; Saito, H.; Kouyama, Y.; Mimori, K.; Ando, T.; Watanabe, S.;
Oshima, M., Nano-scale physical properties characteristic to metastatic intestinal cancer cells identified by high-speed scanning ion
conductance microscope. Biomaterials 2022, 280, 121256
Watanabe, S.; Kitazawa, S.; Sun, L.; Kodera, N.; Ando, T.; Development of High-Speed Ion Conductance Microscopy, Rev. Sci. Instrum.
2019, 90(12), 123704.

31

Takeshi Fukuma

http://fukuma.w3.kanazawa-u.ac.jp/

Visualizing Nanoscale Dynamics andMechanics in Living Cells by Nanoendoscopy AFM

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

Takeshi Fukuma

E-mail: fukuma@staff.kanazawa-u.ac.jp
Website: http://fukuma.w3.kanazawa-u.ac.jp/

Research Interests
Atomic Force Microscopy, Nanoscale Measurement Technologies, Electrical Engineering, Interfacial Sciences, Life Sciences, Electrochemistry

Education
2003

Doctor of Engineering, Department of Electronic Science and Engineering, Kyoto University

Professional Experience2017 – present2017 – present
2012 – 20172007 – 20122005 – 2007
2001 – 2005

Director/Professor, Nano Life Science Institute (WPI-NanoLSI), Kanazawa University
Professor, Faculty of Frontier Engineering, Kanazawa University
Professor, Faculty of Electronic Eng. and Computer Sci., Kanazawa University
Associate Professor, Frontier Science Organization, Kanazawa University
Senior Scientist, Physics Department, Trinity College Dublin (Ireland)
Research Fellow, Kyoto University

Honors
2023
2018
2017
2011

Science and Technology Award (Research), The Commendation for Science and Technology by MEXT
15th JSPS Prize, Japan Society for the Promotion of Science

Atomic force microscopy (AFM) is the only method that allows direct imaging of nanoscale dynamics
of biomolecules in liquid without labeling. However, such high-resolution imaging is possible only forbiological systems reconstructed on a solid substrate. To overcome this limitation, we have developed
nanoendoscopy AFM, where we insert a needle-like probe into a living cell and perform AFM
measurements of the intra-cellular nanoscale dynamics and mechanics. This technique has two
distinctive advantages over existing bio-imaging tools. First, it allows high-resolution imaging at aninterface, such as the inner surface of the bottom cell membrane or the upper surface of the nucleus.
To take advantage of this feature, we are investigating the structure and dynamics of focal adhesions(FAs). FAs connect actin fibers and extra-cellular structures and play important roles in cell motility
and adhesion. Combining nanoendoscopy AFM with a confocal fluorescence microscope, we
succeeded in direct imaging of the growth of a FA and the actin stress fiber associated with it. Such
imaging allows correlating molecular transport visualized by fluorescence microscopy and nanoscale
structural changes observed by AFM, helping us to understand the molecular mechanisms of various
intra-cellular phenomena. Another advantage is the capability of nanomechanical property
measurements. Nanoendoscopy AFM allows direct indentation of an intra-cellular component with aneedle probe and quantitative estimation of local elasticity. With this technique, we are investigating
nuclear elasticity changes induced by metastatic cancer progression. While the cell membranes
become softer with the cancer progression, the nuclear membranes become stiffer. Our preliminary
biochemical analyses suggest that the stiffening is due to increased chromatin compaction levels.
Such a stiffening may imply its biological role in protecting the DNA from the risk of nuclear rapture
during their invasive migration, where high pressure will be applied to the nucleus in passing througha small channel.

Hokkoku Bunka Award, Hokkoku Shinbun
Young Scientists’ Prize, The Commendation for Science and Technology by MEXT

Publications

32

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

1.

Yurtsever, A.; Wang, P. X.; Priante, F.; Morais Jaques, Y.; Miyazawa, K.; MacLachlan, M. J.; Foster, A. S.; Fukuma, T., Molecular insights
on the crystalline cellulose-water interfaces via three-dimensional atomic force microscopy. Sci. Adv. 2022, 8 (41), eabq0160.

2.

Yurtsever, A.; Wang, P. X.; Priante, F.; Morais Jaques, Y.; Miyata, K.; MacLachlan, M. J.; Foster, A. S.; Fukuma, T., Probing the Structural
Details of Chitin Nanocrystal–Water Interfaces by Three ‐ Dimensional Atomic Force Microscopy. Small Methods 2022, 6, 2200320.

3.

Penedo, M.; Miyazawa, K.; Okano, N.; Furusho, H.; Ichikawa, T.; Alam Mohammad, S.; Miyata, K.; Nakamura, C.; Fukuma, T.,
Visualizing intracellular nanostructures of living cells by nanoendoscopy-AFM. Sci. Adv. 2021, 7 (52), eabj4990.

4.

Fukuma, T.; Garcia, R., Atomic- and Molecular-Resolution Mapping of Solid-Liquid Interfaces by 3D Atomic Force Microscopy. ACS
Nano 2018, 12 (12), 11785-11797.

References1.

Penedo, M.; Miyazawa, K.; Okano, N.; Furusho, H.; Ichikawa, T.; Alam Mohammad, S.; Miyata, K.; Nakamura, C.; Fukuma, T., Visualizing
intracellular nanostructures of living cells by nanoendoscopy-AFM. Sci. Adv. 2021, 7 (52), eabj4990.

2.

Penedo, M.; Shirokawa, T.; Alam, M. S.; Miyazawa, K.; Ichikawa, T.; Okano, N.; Furusho, H.; Nakamura, C.; Fukuma, T., Cell penetration
efficiency analysis of different atomic force microscopy nanoneedles into living cells. Sci. Rep. 2021, 11 (1), 7756.

33

Salvatore Chiantia

https://www.uni-potsdam.de/en/ibb-physbiochem/index

Professor, Doctor
Inst. of Biochemistry and Biology, University of Potsdam, Germany

E-mail: chiantia@uni-potsdam.de
Website: https://www.uni-potsdam.de/en/ibb-physbiochem/index

Research Interests
Fluorescence microscopy, fluctuation analysis, virus assembly, membrane biophysics

Education
2008

Doctor of Physics, University of Dresden, Germany

Professional Experience
2021 – present
2015 – 2021
2014 – 20152012 – 20142009 – 20122008 – 2009

Professor of Physical Biochemistry, University of Potsdam, Germany
Junior Professor of Physical Cell Biochemistry, University of Potsdam, Germany
Principal Investigator, Dept. of Biology, Humboldt University, Berlin, Germany

Investigating Virus-host Interactions
at a Single Cell Level Using Quantitative
Fluorescence Microscopy
Salvatore Chiantia
Inst. of Biochemistry and Biology, University of Potsdam, Germany

The pathogenesis of influenza A viruses (IAVs) is influenced by several factors, including IAV strain
origin and reassortment, tissue tropism and host type. While such factors were mostly investigated in
the context of virus entry, fusion and replication, little is known about the viral-induced changes to thehost lipid membranes during assembly. In this work, we applied several biophysical fluorescencemicroscope techniques (i.e., membrane charge Förster energy resonance transfer sensing,
generalized polarization imaging and scanning fluorescence correlation spectroscopy (sFCS)) toquantify the effect of infection by two IAV strains of different origin (avian: FPV; human: WSN) in
avian and human cell lines (DF1 and HEK293T). We found that IAV infection affects the membranecharge of the inner leaflet of the plasma membrane. Moreover, we showed that IAV infection impacts
lipid-lipid interactions by decreasing membrane fluidity and increasing lipid packing. Because of suchalterations, diffusive dynamics of membrane-associated proteins were hindered. Taken together, ourresults indicate that the infection of avian and human cell lines with IAV strains of different origins had
similar effects on the biophysical properties of the plasma membrane at the assembly side.

Postdoc, Dept. of Biology, Humboldt University, Berlin, Germany
Postdoc, Dept. of Biochemistry and Cell Biology, State University of New York, USA
Postdoc, Dept. of Physics, Technical University of Dresden, Germany

Honors
2009
2008
2007

LSRF Fellowship, sponsor Howard Hughes Medical Institute (HHMI)
Feodor Lynen Fellowship, Alexander von Humboldt Foundation
Student Research Achievement Award, Biophysical Society

Publications

34

1.

Dunsing V, Petrich A, Chiantia S. Multicolor fluorescence fluctuation spectroscopy in living cells via spectral detection- eLife 2021, doi:
10.7554/eLife.69687

2.

Petrich A, Dunsing V, Bobone S, Chiantia S. Influenza A M2 recruits M1 to the plasma membrane: A fluorescence fluctuation
microscopy study. Biophysical Journal 2021, doi: 10.1016/j.bpj.2021.11.023

3.

Petazzi RA, Aji AK, Tischler ND, Chiantia S. Detection of Envelope Glycoprotein Assembly from Old-World Hantaviruses in the Golgi
Apparatus of Living Cells. Journal of Virology 2020, JVI.01238-20.

35Daniel J. Müller

Mechanically Quantifying
and Guiding Biological Processes

https://bsse.ethz.ch/biophysics

Professor of Biophysics, Doctor

Daniel J. Müller

Department of Biosystems Science and Engineering, ETH Zürich, Switzerland

ETH Zurich, Biosystems Science and Engineering, Basel, Switzerland

E-mail: daniel.mueller@bsse.ethz.ch
Website: https://bsse.ethz.ch/biophysics

Research Interests
Biophysics, Bionanotechnology, Membrane Proteins, Atomic Force Microscopy, Cellular Systems

Education
1997

PhD in Biophysics, Biozentrum, University of Basel, Switzerland

Professional Experience
2021 – present
2020 – present
2019 – 2021
2014 – present2010 – present
2003 – 2005
2002 – 2010
2000 – 2002

Head of the Department of Biosystems Science and Engineering, ETH Zürich, Switzerland.
Foreign Scientific Member of the Max-Planck-Institute of Medical Research, Heidelberg, Germany.
Co-Head of the Department of Biosystems Science and Engineering, ETH Zürich, Switzerland.
Co-Director of the NCCR Molecular Systems Engineering, Switzerland.
Chair of Biophysics, Department of Biosystems Science and Engineering, ETH Zürich, Switzerland.
Director of the Biotechnology Center, Dresden, Germany.
Full professor for Cellular Machines (C4-position) at the Biotechnology Center, University of Technology Dresden, Germany
Groupleader (C3-position) at the Max-Planck-Institute of Molecular Cell Biology and Genetics, Dresden, Germany..

Honors
2023
2022
2020
2019
2016
2011

Mechanobiology emerges at the crossroads of medicine, biology, biophysics and engineering anddescribes how the response of proteins, cells, tissues and organs to mechanical cues contribute todevelopment, differentiation, physiology and disease. The grand challenge in mechanobiology is toquantify how biological systems sense, transduce, respond and apply mechanical signals. Over three
decades, atomic force microscopy (AFM) has emerged as a key platform enabling the simultaneous
morphological and mechanical characterization of living biological systems. Here, we will introduce
the use of AFM-based nanoscopic assays to mechanically characterize, stimulate and animal cells
and cellular systems. For example, we will exemplify how to use AFM-based assays to characterize
viruses binding to mammalian cells and demonstrate how to use these insights to direct virus infection
in vitro and in vivo for controlling cellular function and to restore vision. In another example we will
use AFM to characterize how adherent cells sense the extracellular matrix (ECM) and responds to its
biochemical composition and mechanical properties. Finally, we will employ AFM to functionallystimulate neuronal networks while following their electrophysiological response and to understandhow individual neurons sense and respond to mechanical stimuli.

Honorary Professor of the University of Heidelberg, Germany

References
1.
2.
3.
4.
5.
6.

Atomic force microscopy imaging modalities in molecular and cell biology. Y.F. Dufrêne et al., Nature Nanotechnology (2017) 3, 295.
Atomic force microscopy-based characterization and design of biointerfaces. D. Alsteens et al., Nature Review Materials (2017) 2, 17008.
Combined activities of hydrostatic pressure and the actomyosin cortex drive mitotic cell rounding. M.P. Stewart et al., Nature (2011) 469, 226.
Nanomechanical mapping of first binding steps of a virus to animal cells. D. Alsteens et al., Nature Nanotechnology (2017) 12, 177.
Virus stamping for targeted single cell infection in vitro and in vivo. R. Schubert et al., Nature Biotechnology (2018) 36, 85.
AFM-based Mechanobiology. M. Krieg et al., Nature Reviews Physics (2019).

International Research Award of Germany, Humboldt Foundation, Ministry of Science and Education (BMBF), Germany
Scientific Member of Max-Planck Society, Germany
Marsilius Medal of the University of Heidelberg, Germany
Elected EMBO Member
Fellow of the Center of Advanced Studies of the Ludwig-Maximillian University, Munich, Germany

Publications

36

1.

In mitosis integrins reduce adhesion to extracellular matrix and strengthen adhesion to adjacent cells
M. Huber, J.C. Casares-Arias, R. Fässler, D.J. Müller & N. Strohmeyer. Nature Communications (2023) 14, 2143

2.

Gasdermin-A3 pore formation propagates along variable pathways
S.A. Mariet al. Nature Communications (2022) 13, 2609.

3.

The nucleus acts as a ruler tailoring cell responses to spatial constraints
A.J. Lomakin, et al. Science (2020) 370, eaba2894.

4.

Protease-activated receptors use multiple pathways to activate α5β1 integrins and assemble adhesomes in non-hematopoietic cells
P.M. Spoerri, N. Strohmeyer, Z. Sun, R. Fässler & D.J. Müller. Nature Materials (2020) 19, 218–226

5.

Atomic force microscopy-based mechanobiology
M. Krieg, et al. Nature Reviews Physics (2019) 1, 41-57.
.
37

Tomoyoshi Yamano

Development of Engineered Exosome
for Immune Regulation

http://immunology.w3.kanazawau.ac.jp/index2.html

Associate Professor, Doctor
Graduate School of Medical Sciences, Kanazawa University, Japan
Nano Life Science Institute (WPI-NanoLSI), Kanazawa University, Japan

Tomoyoshi Yamano

E-mail: tomoyoshi.yamano@med.kanazawa-u.ac.jp
Website: http://immunology.w3.kanazawa-u.ac.jp/index2.html

Research Interests
Immunology, Exosome Engineering, Synthetic biology

Education
2011

Division of Immunobiology, Institute for Biological Sciences, Tokyo University of Science, Japan.

Professional Experience
2023 – Present
2022 – Present
2019 – 2023
2017 – Present2017 – 2022
2011 – 20172011 – 2011

JST FOREST Researcher
Associate professor in Graduate School of Medical Sciences, Kanazawa University, JapanJST PRESTO Researcher
Associate professor in Nano Life Science Institute (WPI-NanoLSI), Kanazawa University, Japan

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

Exosomes play pivotal roles in cell-cell communication. It has been demonstrated that any protein ofinterest can be expressed on exosomes through conjugation with tetraspanins, exosome-specificsurface proteins. Leveraging this method, we have developed engineered exosomes thatsimultaneously express several immune modulators. A distinctive feature of this approach is that target
cells can concurrently receive all signals from these immune modulators. Using our method, we havedeveloped several engineered antigen-presenting exosomes capable of either positively or
negatively regulating immune responses. For example, Antigen Presenting Exosomes (AP-Exo-CTL)
are engineered to express a peptide-major histocompatibility class I (pMHCI) complex, acostimulatory CD80 molecule, and IL-2 on their surface, thus enabling the simultaneous presentationof multiple immune modulators to antigen-specific CD8 T cells. This leads to the clonal expansion anddifferentiation of antigen-specific CTL, inducing potent anti-cancer effects. Conversely, AP-Exo-Treg
exosomes are engineered to express the antigen-MHCII complex, IL-2, and TGF-β on their surface,promoting the induction of antigen-specific regulatory T cells. We will discuss the potential andefficacy of these engineered exosomes in modulating immune responses.

Assistant professor in Department of Immunology, Kanazawa University, Japan
Postdoctoral researcher in Institute for Immunology, Ludwig-Maximilians University, Germany
Visiting researcher in Department of Dermatology, University Hospital Zurich, Switzerland

References
1.

Publications

38

1.

Keesiang Lim, Goro Nishide, Elma Sakinatus Sajidah, Tomoyoshi Yamano, Yujia Qiu, Takeshi Yoshida, Akiko Kobayashi, Masaharu
Hazawa, Toshio Ando, Rikinari Hanayama, Richard W Wong., Nanoscopic Assessment of Anti-SARS-CoV-2 Spike Neutralizing
Antibody Using High-Speed AFM. Nano letters. 2023, 23(2) 619-628.

2.

Xiabing Lyu, Shota Imai, Tomoyoshi Yamano, Rikinari Hanayama., Preventing SARS-CoV-2 Infection Using Anti-spike
Nanobody-IFN-β Conjugated Exosomes. Pharmaceutical research, 2022, 40(4) 927-935

3.

Yamano T, Dobeš J, Vobořil M, Steinert M, Brabec T, Ziętara N, Dobešová M, Ohnmacht C, Laan M, Peterson P, Benes V, Sedláček R,
Hanayama R, Kolář M, Klein L, Filipp D., Aire-expressing ILC3-like cells in the lymph node display potent APC features. The Journal ofexperimental medicine. 2019, 216(5) 1027-1037

2.
3.

Keesiang Lim, Goro Nishide, Elma Sakinatus Sajidah, Tomoyoshi Yamano, Yujia Qiu, Takeshi Yoshida, Akiko Kobayashi, Masaharu
Hazawa, Toshio Ando, Rikinari Hanayama, Richard W Wong., Nanoscopic Assessment of Anti-SARS-CoV-2 Spike Neutralizing Antibody
Using High-Speed AFM. Nano letters. 2023, 23(2) 619-628.
Xiabing Lyu, Shota Imai, Tomoyoshi Yamano, Rikinari Hanayama., Preventing SARS-CoV-2 Infection Using Anti-spike Nanobody-IFN-β
Conjugated Exosomes. Pharmaceutical research, 2022, 40(4) 927-935
Yamano T, Dobeš J, Vobořil M, Steinert M, Brabec T, Ziętara N, Dobešová M, Ohnmacht C, Laan M, Peterson P, Benes V, Sedláček R,
Hanayama R, Kolář M, Klein L, Filipp D., Aire-expressing ILC3-like cells in the lymph node display potent APC features. The Journal ofexperimental medicine. 2019, 216(5) 1027-1037

39

POSTER SESSIONS

40

41

P-1

P-2

The 7th NanoLSI Symposium in Berlin; From Molecules to Cells and Tissue – Bridging Scales with Nanoprobe
Technology
Nanoscale visualization of Aptamer-CYP24 complex binding dynamics in the development of aptamer
therapeutics in cancer

Light-induced manipulation of colloidal particles in a photosensitive polymer-dye system
López de Guereñu A., Lomadze N., Santer S.
University of Potsdam, Faculty of Physics and Astronomy

Madhu Biyani
WPI Nano Life Science Institute (WPI-NanoLSI), Kanazawa University, Kanazawa, Japan.
biyani@staff.kanazawa-u.ac.jp
Abstract
Overexpression of a mitochondrial enzyme CYP24 in cancer cells decreases the level of vitamin D3, which
has anti-cancer activity [1]. To enhance vitamin D3 functionality in cancer cells, we identified a CYP24inhibiting DNA aptamer (Apt-7). In this study, we used high-speed atomic force microscopy (HS-AFM) [2] andmolecular docking methods to provide a single-molecule explanation of the aptamer-based CYP24 inhibition
mechanism [3]. The HS-AFM revealed the dynamic interactions of Apt-7 and CYP24 at the nanoscale level andthe spatio-temporal analysis of captured images enabled us to characterize the binding interaction of Apt-7 andCYP24. The molecular docking results further revealed that Apt-7 combined steadily at active or allostericbinding sites of CYP24 by strong hydrogen bonds and electrostatic interaction. Interestingly, images from HSAFM showed remarkably similar conformation with molecular docking results. In summary, the integrated
approach of HS-AFM and molecular docking provided a bridge to the development of a lead molecule forpromising anticancer therapy.

e-mail: lopezdeguerenukurganova@uni-potsdam.de
We present a multi-dimensional investigation into the optical and physical characteristics of a complex
photosensitive surfactant system. This system comprises mesoporous silica particles in an aqueous solution of
a fluorescent dye and an azobenzene-containing surfactant, capable of binding into a complex together. The
azobenzene-containing surfactant undergoes reversible photo-isomerization from its trans- to cisconfiguration upon exposure to UV/blue light, with this isomerization process occurring on a picosecond
timescale. This unique property enables precise control over the dynamic aggregation or dispersion ofmesoporous colloidal particles upon illumination. The formation of the dye/surfactant complex introduces anadditional dimension to the investigation of the described system.
The driving force behind the possibility of remotely controlling the colloids is the phenomenon of light-driven
diffusioosmosis (LDDO). The porous structure of mesoporous silica colloids allows them to act as a source orsink of the photosensitive surfactant, driving the “local” LDDO process and leading to mutual interactions(repulsion/attraction) between particles, which may be used to impose a fine structure on a 2D particle grid.
Depending on the isomerization state, the surfactant has different adsorption affinities to both the particles andthe fluorescent dye. The trans-isomer readily enters and accumulates in the negatively charged pores of theparticles, while also forming a complex with the negatively charged organic dye. However, the cis-isomer isexpulsed out of the pores and remains in the solution outside the particles; it also doesn’t bind to the dye. Thus,
upon irradiation, the azobenzene-containing surfactant isomerizes to its cis-state within the colloids’ pores, thedye-surfactant complex gets destroyed and the polymer readily diffuses out, generating a decrease inluminescence and an excess concentration near the colloid’s outer surface, which leads to the creation of adiffusioosmotic flow and the displacement of the particles. This process is wavelength-and intensitydependent, allowing for the control over these structures. The dye provides an additional dimension to thesystem, adding to its complexity due to its influence on the surfactant isomerization and the LDDO process.
References:
1. Feldmann, D.; Arya, P.; Molotilin, T. Y.; Lomadze, N.; Kopyshev, A.; Vinogradova, O. I.; Santer, S.
Extremely Long-Range Light-Driven Repulsion of Porous Microparticles. Langmuir (2020).
2. Feldmann, D.; Maduar S.R.; Santer, M.; Lomadze, N.; Vinogradova O.I.; Santer, S. “Manipulation ofsmall particles at solid liquid interface: light driven diffusioosmosis”. Scientific Reports, (2016).
3. Arya P.; Feldmann, D.; Kopyshev, A.; Lomadze N.; Santer, S. Light driven guided and self-organizedmotion of mesoporous colloidal particles. Soft Matter, (2020).
4. Sharma, A.; Jung, S-H.; Lomadze, N.; Pich, A.; Santer, S.; Bekir, M. “Adsorption Kinetics of a
Photosensitive Surfactant Inside Microgels.” Macromolecules, (2021).
5. Arya, P.; Jelken, J.; Feldmann, D.; Lomadze, N.; Santer, S. Light driven diffusioosmotic repulsion andattraction of colloidal particles. J. Chem. Phys. (2020).

References:
1.
2.
3.

42

Deeb, K., Trump, D. & Johnson, C. Vitamin D signaling pathways in cancer: potential for anticancer
therapeutics. Nat Rev Cancer 2007, 7, 684–700.
Toshio Ando, Takayuki Uchihashi, and Simon Scheuring, Chemical Reviews 2014, 114, 6, 3120-3188.
Madhu Biyani, Kaori Yasuda, Yasuhiro Isogai, Yuki Okamoto, Wei Weilin, Noriyuki Kodera, Holger
Flechsig, Toshiyuki Sakaki, Miki Nakajima, and Manish Biyani, ACS Applied Materials & Interfaces
2022, 14, 16, 18064-18078.

43

P-3

P-4

Nanoscopic Evaluation of Anti-SARS-CoV-2 Spike
Neutralizing Antibody Using High-Speed AFM

Local direction of opto-mechanical stress in azobenzene containing
polymers during surface relief grating formation
Sarah Loebner,1 Bharti Yadav,2 Nino Lomadze,1 Nina Tverdokhleb,2Hendrik Donner,3 Marina Saphiannikova,2 Svetlana Santer1

Lim Kee Siang

WPI Nano Life Science Institute, Kanazawa University

1

Institute of Physics and Astronomy, University of Potsdam, 14476 Potsdam, Germany
2

Leibniz Institute of Polymer Research Dresden, 01069 Dresden, Germany
CADFEM GmbH, 09121 Chemnitz, Germany

3

Abstract: SARS-CoV-2, thenewly emerging ȕ-coronavirus,
has caused the unprecedented
COVID-19 pandemic since 2019.
Delayed onset of COVID-19 plus
highly infectious variants haveaccelerated the infection rate toworldwide. Although several
mRNA vaccines against viral
spike (S) protein have beendeveloped to prevent the disease,the protection wanes fast andoverride by new variants. Anti-S
protein neutralizing antibodies (SNAbs) provide an effective
alternative to prevent and treat
COVID-19, in which people canconstantly receive rapid passive
immunity against SARS-CoV-2.
Direct visualization of the
dynamic S NAb and S protein
interaction at nanoscopic level
using current imaging tools, cryoEM and confocal imaging forexample, remains difficult. We
here perform high-speed atomic
force microscopy (HS-AFM) imaging to elucidate this interaction, in which it is essential to study the binding
pattern of S NAbs and the risk of antigen-dependent enhancement (ADE). We first observed the innate
structural properties of an S NAb (MM43; IgG1) such as the Y conformation of IgG antibody, and formationof spontaneous hexameric rings at high density. The S NAb neither induce RBD opening nor S1 subunit
shedding after it bound to S protein. On the other hand, we noticed that the S NAb-S protein interaction was
stable at acidic pH (pH5). Collectively, these findings suggested that the S NAb has low risk of ADE. Besides
protein-protein interaction, we also developed small extracellular vesicles with surface S protein (S sEV) to
mimic the S protein of SARS-CoV-2. Dynamic interaction between S NAb and S sEV together with thebinding tendency of S NAb could be evaluated through HS-AFM scanning. In summary, we demonstrate ananoscopic platform that is feasible to study the binding properties of numerous S NAbs either isolated fromconvalescent COVID-19 patients or via antibody design.

Contact: loebner@uni-potsdam.deOn this poster we present how the photo-induced deformation of azobenzene containing polymers relates
to the local direction of opto-mechanical stresses generated during irradiation with interference patterns(IPs). We can substantiate the modeling approach proposed in Ref. 1, which describes the directional photodeformations in glassy side-chain azobenzene polymers, and prove that these deformations arise from there-orientation of rigid backbone segments along the light polarization direction. In experiments we inscribe
surface relief grating in pre-elongated photosensitive colloids of few micrometers length using different IPs
such as SS, PP, +45, SP, RL and LR. The deformation of the colloidal particles is studied in-situ, whereby the
local variation of the polymer topography is assigned to the local distribution of electrical field vector for all
IPs. Orientation approach correctly predicts local variations of the main axis of light-induced stress in eachinterference pattern for both initially isotropic and highly oriented materials. With this work, we suggest thatthe orientation approach implements a self-sufficient and convincing mechanism to describe photo-induced
deformation in azobenzene containing polymer films that in principle does not require auxiliary assumptions.

Figure 1. AFM micrographs showing SRG inscribed in the azobenzene containing polymer film using different
interference pattern. White dashed horizontal lines indicate positions at the micrographs where the IP isswitched. Additionally, the distribution of the electrical field vectors (white arrows) are assigned to the
topography variation.
References:[1] Yadav, B.; Saphiannikova, M.; Domurath, J.; Kim, K.; Lee, S. Orientation approach to directional
photodeformations in glassy side-chain azopolymers. J. Phys. Chem. B 2019, 123, 3337-3347.

44

45

P-5

P-6

Deciphering actin structure-dependent preferential cooperative binding of cofilin
Kien Xuan Ngo1, Huong T Vu2, Kenichi Umeda1, Noriyuki Kodera1, Taro Q.P. Uyeda3, Toshio Ando1
Nano Life Science Institute (WPI-NanoLSI), Kanazawa University, Kanazawa, Ishikawa 920-1192, Japan.
Centre for Mechanochemical Cell Biology, Warwick Medical School, Coventry CV4 7AL, UK.
3
Department of Physics, Faculty of Science and Engineering, Graduate School of Waseda University, Shinjuku, Tokyo 1698555, Japan.
Corresponding Email: ngoxuakien@staff.kanazawa-u.ac.jp
1

Light-Triggered Manipulations of Droplets All in One:
Reversible Wetting, Transport, Splitting, and Merging

2

In our earlier study, we observed that the shortened helical pitch in cofilactin, where a cofilincluster is bound to an actin filament, affected the neighboring bare zone to shorten a half helical pitch(HHP) on the pointed-end (PE) side of the cluster, while the pitch on the barbed-end (BE) side remained
similar to the control. The unidirectional growth of cofilin clusters toward the PE of the filamentsuggested that cofilin favors actin structures with a shorter HHP. However, the exact mechanism
underlying the cooperative binding of cofilin to actin filaments based on their favorable actin structure
has remained unclear. To delve into this mechanism, we conducted a principal component analysis on
actin structures derived from 46 PDB structures. This analysis classified the structural differences in actin
associated with different nucleotides and actin binding proteins. In particular, we found a notable contrast
in the structure of ADP-actin between F-actin and cofilactin, providing a structural basis for suggestingthat the influence of ADP has a minimal effect on actin structure that is conducive to the preferential
binding of cofilin. Furthermore, through the use of high-speed atomic force microscopy, we made a novel
discovery: the bare half helix with shortened HHP on the PE side of the cluster comprised fewer actin
protomers compared to the normal helices. The mean axial distance between two adjacent actin
protomers along the same long-pitch strand, referred to as MAD, in the shortened bare HHP exhibited
elongation and thermal fluctuation in a higher range (5.0 – 6.3 nm) than the MAD in the normal HHP(4.3 – 5.6 nm). The impediment of cooperative structural changes in helical twisting, achieved throughstronger attachment to the lipid membrane, had a more pronounced hindrance on cofilin binding and the
expansion of larger clusters than the impact of inorganic phosphate. Collectively, we hypothesize thatthe shortening of half helices, resulting in elongation in MAD, assumes a pivotal role in facilitating thepreferential cooperative binding of cofilin to actin protomers.

Maren Umlandt, Alexey Kopyshev, Sergey V. Pasechnik, Alexandre V. Zakharov, Nino Lomadze,
Svetlana Santer,
Smart Soft Matter, Institute of Physics and Astronomy, University of Potsdam, Germany
Contact: mrabe@uni-potsdam.deHere we report on light-triggered droplet manipulation such as reversible wetting, splitting, merging, andtransport [1]. The unique feature of our approach is that the changes in the wetting properties of
microscopic droplets of anisotropic (liquid crystalline) liquids adsorbed on photo-switchable films canbe triggered just by application of soft optical stimuli, which lead to dynamical, reversible changes in thelocal morphology of the structure surfaces. The adaptive films consist of an azobenzene-containing
surfactant ionically attached to oppositely charged polymer chains. Under exposure to irradiation withlight, the azobenzene photo-isomerizes between two states, nonpolar trans- and polar cis-isomers,
resulting in corresponding changes in surface energy and orientation of the surfactant tails at the interface[2]. Additionally, the local increase in the surface temperature due to absorption of light by theazobenzene groups [3] enables diverse processes of manipulation of the adsorbed small droplets, such as
the reversible increase of the droplet basal area up to 5 times, anisotropic wetting during irradiation withmodulated light, and precise partition of the droplet into many small pieces. Moreover, using a moving
focused light spot, we demonstrate the locomotion of the droplet over macroscopic distances with avelocity of up to 150µm∙s-1. Our findings could lead to the ultimate application of a programmable
workbench for manipulating and operating an ensemble of droplets, just using simple and gentle opticalstimuli.
[1] Umlandt, M.; Kopyshev, A., Pasechnik, S.V., Zakharov, A.V., Lomadze, N., Santer, S. LightTriggered Manipulations of Droplets All in One: Reversible Wetting, Transport, Splitting, and Merging
ACS Applied Materials & Interfaces 2022 14 (36), 41412-41420 DOI: 10.1021/acsami.2c10710[2] Arya, P.; Jelken, J.; Lomadze, N.; Santer, S.; Bekir, M. Kinetics of Photo-isomerization of
Azobenzene Containing Surfactants. J. Chem. Phys. 2020, 152, 024904 DOI: 10.1063/1.5135913[3] Zakharov, A. V.; Maslennikov, P. V. Laser-excited Motion of Liquid Crystals Confined in a
Microsized Volume with a Free Surface. Phys. Rev. E 2017, 96, 052705 DOI:
10.1103/PhysRevE.96.052705

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P-8

Real-time visualization of Structural Maintenance of Chromosomes complexes byhigh-speed atomic force microscopy

Characterization and optimization of SERS substrates using
adenine as benchmark molecule and automation of data evaluation

Kenichi Umeda

Vı́t Pavelka1,∗ , Dušan Hemzal2 , Hitoshi Asakawa3 , Naoki Kida3 and Jan Hrbáč1Nano Life Science Institute (WPI-NanoLSI), Kanazawa University
umeda.k@staff.kanazawa-u.ac.jp

1

Department of Chemistry, Masaryk University, Brno, CZ
Department of Condensed Matter Physics, Masaryk University, Brno, CZ
3Faculty of Chemistry, Institute of Science and Engineering, Kanazawa University, Kanazawa, Japan∗
Corresponding author: Vı́t Pavelka, 408930@mail.muni.cz, pavelka.vit69@gmail.com
2

Many of chromosomal functions are linked to the folding structure of genome DNA, which is meters in length but
packed in a micrometer-order nucleus. The structural maintenance of chromosomes (SMC) complexes, including
cohesin, condensin, and Smc5/6, play central roles in such three-dimensional genome organization. SMC
complexes are ring-shaped ATPase assemblies that topologically encircle DNA strand inside of their ring to form
physical links between distantly separated DNA segments. Resent biochemical and single-molecule studies haverevealed dynamic behaviors of SMC complexes, however, it remains elusive how the proteinous rings bind andmanipulate DNA at the sub-molecular level. In this study, we applied high-speed AFM to directly visualize SMC
complexes that bound to DNA. This visualized the structural changes of single SMC molecule with their various
behavers on DNA, like one-dimensional diffusion, at 100 millisecond-order resolution. Furthermore, our results
obtained with the ATPase mutant suggested that SMC complex initially associates with DNA via ATPase domain,which follows to the late DNA-binding state. Based on these results, we discuss the molecular mechanism bywhich SMC complexes bind to DNA.

Abstract
Providing insight into biomolecular interactions at the nanoscale, Surface-Enhanced Raman
Spectroscopy (SERS) is an important tool in biophysics and structural chemistry [1]. Factors
such as morphology, shape, and type of nanostructured metal, laser wavelength, and analyte
choice affect the performance of SERS substrates. Consequently, efforts invested in characterizing and improving these substrates to enhance their sensitivity, reproducibility, uniformity,
and affordability, are paramount.
While the conventional use of fluorescent dyes for benchmarking of the SERS substrates
introduces complexities due to resonance Raman scattering effects [2], we focus on adenine asan optimal reporting molecule for characterization of the SERS substrates. Adenine exhibits
several advantageous properties making it an ideal choice, particularly for silver substrates.
Importantly, adenine is non-resonant at visible wavelengths, but possesses strong Raman crosssection. Furthermore, in the 1 µM − 10 µM concentration range a unique surface complex
between silver and adenine forms, providing additional Raman enhancement and presenting
opportunities for biophysical investigations [3]. At higher adenine concentrations, we evidenced
a layer of oriented adenine molecules on the surface of silver SERS substrates by atomicallyresolved 3D AFM.
The large-area SERS substrates characterization and optimization generally represent a demanding task, that requires evaluation of vast Raman datasets. Therefore, we present a novel
approach enabling a precise automated SERS background removal and peak identification
based on morphology operators [4].

References[1] Sebastian Schlücker. Surface Enhanced Raman Spectroscopy: Analytical, Biophysical andLife Science Applications. John Wiley & Sons, Ltd, 2010.
[2] Claudia Fasolato. Surface Enhanced Raman Spectroscopy for Biophysical Applications:
Using Plasmonic Nanoparticle Assemblies. Springer Theses. Springer International Publishing, 2018.
[3] Evanthia Papadopoulou and Steven E. J. Bell. Surface enhanced raman evidence for Ag+
complexes of adenine, deoxyadenosine and 5’-dAMP formed in silver colloids. Analyst,
135(12):3034–3037, 2010. Publisher: The Royal Society of Chemistry.
[4] Vı́t Pavelka, Dušan Hemzal, and Jan Hrbáč. Complex evaluation of raman spectra usingmorphological filtering: Algorithms, software implementation, and experimental verification of baseline correction, peak recognition, and cosmic ray removal in SERS spectra ofdesigner drugs. Journal of Raman Spectroscopy, 53(12):2100–2109, 2022.

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P - 10High-speed atomic force microscopy revealed alternative interactions between CaMKII holoenzymes
depend on their phosphorylation

Exploring the mechanics of cortical dendrite formation through AFM measurements

Taisei Suzuki1, Hideji Murakoshi2, Mikihiro Shibata3,4
1Grad. Sch. NanoLS., Kanazawa Univ.

Rei Yagasaki, Satoru Okuda

2Supportive Center for Brain Research., NIPS

Nano Life Science Institute, Kanazawa University

3WPI-Nano Life Science Institute, Kanazawa Univ.

Contact: rei-yagasaki@staff.kanazawa-u.ac.jp

4Institute for Frontier Science Initiative, Kanazawa Univ.

Contact: Taisei Suzuki

Abstract
Dendrites are important structures that receive signals from other neurons. Previous studies
have shown that dendrites elongate and branch in response to the signals. Recently,
mechanical properties have attracted much attention as a new factor in regulating cell
behavior, and it is thought that they also affect dendrite elongation and branching. However,
the mechanical properties of dendrites and the surrounding environment are not wellunderstood, and the effect of the mechanical properties on dendrites is unknown. Therefore,
to investigate the influence of mechanical properties on dendrites, we isolated and cultured
neurons from mouse cortex and measured the properties by atomic force microscopy (AFM).
In this presentation, we will report the results obtained so far and discuss future analyses andthe influence of mechanical properties on dendrites. We believe that our work will allow us tounderstand how the mechanical properties of the brain and the nerves themselves influence
the formation of neural networks.

bba0606@stu.kanazawa-u.ac.jp

Long-term potentiation (LTP) and long-term depression (LTD) are considered fundamental cellularlevel mechanisms in memory formation and forgetting, respectively (1), and alter synaptic signal efficiency
and dendritic spine volume (2). At the molecular level, Ca2+/calmodulin-dependent protein kinase II(CaMKII) plays a pivotal role in the functional and structural plasticity of the brain(3). CaMKII is highly
abundant within the post-synaptic density (PSD), comparable to the levels of cytoskeletal proteins (4).
However, its contribution to the mechanism underlying structural plasticity remains elusive. Here, we
employed high-speed atomic force microscopy (HS-AFM) (5), (6), (7) to observe the structural dynamics
of CaMKII at high concentrations similar to those found within the PSD. Furthermore, we observed the
dynamics of the phosphorylation states of CaMKII related to LTP and LTD and analyzed theintermolecular distance between CaMKII holoenzymes. As a result, the interaction among neighboring
CaMKII was dependent on their phosphorylated states. These findings suggest that the structuraldynamics of CaMKII may contribute in part to the structural plasticity in neurons.
References:
1.

T. V. Bliss, T. Lomo, Long-lasting potentiation of synaptic transmission in the dentate area of theanaesthetized rabbit following stimulation of the perforant path. J Physiol 232, 331-356 (1973).

2.

M. Matsuzaki, N. Honkura, G. C. Ellis-Davies, H. Kasai, Structural basis of long-term potentiation in single
dendritic spines. Nature 429, 761-766 (2004).

3.

S. G. Cook, O. R. Buonarati, S. J. Coultrap, K. U. Bayer, CaMKII holoenzyme mechanisms that govern
the LTP versus LTD decision. Sci Adv 7, (2021).

4.

R. Yasuda, Y. Hayashi, J. W. Hell, CaMKII: a central molecular organizer of synaptic plasticity, learningand memory. Nat Rev Neurosci 23, 666-682 (2022).

5.

T. Ando et al., A high-speed atomic force microscope for studying biological macromolecules. Proc Natl
Acad Sci U S A 98, 12468-12472 (2001).

6.

M. Shibata et al., Real-space and real-time dynamics of CRISPR-Cas9 visualized by high-speed atomic
force microscopy. Nat Commun 8, 1430 (2017).

7.

S. Tsujioka et al., Imaging single CaMKII holoenzymes at work by high-speed atomic force microscopy.
Sci Adv 9, eadh1069 (2023).

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P - 12

Measurement of temperature gradients in microscopic volumes using
thermosensitive microgels
Yulia D. Gordievskaya, Nino Lomadze and Svetlana Santer
Institute of Physics and Astronomy, University of Potsdam, 14476 Potsdam, Germany
Contact: yulia.gordievskaya@uni-potsdam.deMicrogels are recognized for their remarkable responsiveness to changes in environmental conditions[1]. In this work, we demonstrated a novel method for local temperature determination in aqueous
solutions by observing the volume-inhomogeneous collapse of microgels positioned near thesubstrate. The substrate consists of a glass cover, a thin titanium oxide layer (<1nm), and a variablethickness gold layer (5-30nm). The localized laser irradiation of the substrate results in the absorptionof light by the gold layer, converting it into heat. We studied how the laser wavelength and power
levels, as well as distance from the irradiation spot influence the size and shape of the thermosensitive
PNIPAM microgel. The obtained dependences allowed us to construct a comprehensive temperature
distribution profile relative to the irradiation point, both along the surface and perpendicular to it.
Moreover, experimental data posed an intriguing question about the underlying mechanisms of heattransfer in liquids. What are the roles of thermophoresis, thermo-osmosis, and convection in thisprocess?

Separation of small extracellular vesicles using an anion exchange
column and revealing their mechanical properties by AFM
Dilireba Bolidong, Takeshi Yoshida, Ayhan Yurtsever, Takayuki Shiba, Takeshi Fukuma, Rikinari
Hanayama
WPI Nano Life Science Institute (NanoLSI), Kanazawa University
Contact: dilireba@staff.kanazawa-u.ac.jp

Abstract
Extracellular vesicles (EVs) are nano-sized vesicles with lipid-bilayer membranes, and secreted byalmost all cell types. Small extracellular vesicles (sEVs) are typically ranging in size from 30 to 150
nanometers in diameter, and they play important roles inintercellular communication, transporting various bioactive
molecules between cells. According to the different biogenesis
mechanisms, EVs are commonly classified into three subtypes
including exosome (EXOs), microvesicles (MVs) and apoptotic
bodies. However, there is currently no method to isolate specificsEVs from a sample containing different types of sEVs, also
their mechanical properties have not been characterized.
In this research, we used an anion exchange column to separate
heterogenous sEVs and revealed the detailed structure andFig 1. Separation of EVs based on anionicchargesand confirmation by EV markers.

nanomechanical properties by Atomic Force Microscopy (AFM). Through the anion-exchange method,sEVs in the culture supernatant can be eluted into different fraction based on surface charge, also
analysis of sEV markers indicated that different anionicFig. A scheme that describes a method for local heating (left). Temperature profile of an aqueous
solution under irradiation with a blue laser in the case of a 30 nm gold layer (right).
[1] Jelken, J., Jung, S. H., Lomadze, N., Gordievskaya, Y. D., Kramarenko, E. Y., Pich, A., & Santer, S.
Tuning the Volume Phase Transition Temperature of Microgels by Light. Advanced Functional Materials,
2022, 32(2), 2107946.

fractions may correspond to different sEV subtypes. (Fig. 1).
Then we isolated the MVs from cells whose plasma membranes
were labeled with a membrane-impermeable biotin reagent anddetected with streptavidin, found that the anion exchange
column can elute MVs into fraction 15 and intraluminal vesicles(ILVs), which are origin of EXOs, into fraction 0 and 12 (Fig.
2). Based on these findings, we separated heterogenous EVs into
MVs and EXOs, then AFM analysis revealed that EXOs haveFig 2. Anionic difference of heterogenous EVs.

higher elasticity and rigidity than MVs. Consequently, wedeveloped a method to clearly separate sEV subtypes derived from different membranes, which
overcomes issues of the current sEV isolation method, also elucidate the structural, mechanicalcharacteristics of heterogenous sEVs.

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P - 14

Reconstruction of Multicellular Patterns with Synthetic Morphogen System

Kosuke Mizuno1,2, Tsuyoshi Hirashima3,4, Satoshi Toda1,2
Graduate School of Frontier Science Initiative, Kanazawa University, Japan
Nano Life Science Institute, Kanazawa University, Japan3
Mechanobiology Institute, National University of Singapore, Singapore
4
Department of Physiology, Yong Loo Lin School of Medicine, National University ofSingapore, Singapore
12Contact: k.mizuno_2010@stu.kanazawa-u.ac.jp

During morphogenesis, cell-cell communication through a secreted factor called morphogen plays
important roles for regulating precise cell position and arrangement. However, the mechanisms through which
diffusing morphogens define precise spatial patterns amidst biological fluctuations remain unclear. Several
theories are reported such as gene regulatory network and endocytic pathway of morphogen, but it is difficult
to separately analyze each factor in model organisms due to the complexity.
Therefore, we designed and developed a SYnthetic Morphogen system for Pattern Logic Exploration using
3D spheroids (SYMPLE3D). This system allowed us to study how cellular responses to concentration
gradients of secreted molecules could generate tissue domains with sharp boundaries. In this system, we
utilized an artificial cell-cell communication through secretion and reception of green fluorescent protein(GFP), using synthetic receptor called synthetic Notch Receptor (synNotch). With this receptor, we canprogram which proteins the cells recognize and which genes the cells turn on in response to that recognition.
We first established GFP-secretor and GFP-receiver cells expressing synNotch. Then, we evaluatedartificial morphogen signaling between these cells after spheroid formation. Here, we explored how the culture
conditions and the expression of responsive genes can influence multicellular patterns in SYMPLE3D.
Interestingly, we found that coupling morphogen signal and cell sorting can sufficiently convert a gradientsignal into distinct tissue domains with a sharp boundary. I will introduce and discuss our findings on patterningin this symposium.
Keywords: Synthetic biology, Morphogen, Cell adhesion

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P - 16Bacterial swimming under flow conditions: fluorescence microscopy, lightdriven micromanipulations and nano-mechanobiology
Valeriia Muraveva1, You-Rong Lin2, Clemens Franz2, Svetlana Santer1, Carsten Beta1
1
Institute of Physics and Astronomy, University of Potsdam, Germany
2
Nano Life Science Institute, Kanazawa University, Japan
Contact: muraveva@uni-potsdam.deClemens M. Franz
Nano Life Science Institute (WPI-NanoLSI), Kanazawa University
Abstract: See page 13

By changing the configuration of the flagella bundle, bacterial swimmers adjust their direction andspeed of locomotion. The soil bacterium Pseudomonas putida, by counterclockwise (CCW) rotationof their flagella, push themselves forward, while by clockwise (CW) rotation they pull themselves in
the opposite direction. Additionally, P. putida can wrap their bundle of flagella around the cellbody.1,2 However, many reasons and benefits of having different motility patterns still remain unclear.
Here, by using different techniques, we study how the swimming pattern of cells changes under flow
conditions.
First, using fluorescence microscopy and microfluidics, we show how the contribution of eachswimming mode changes under laminar shear flow conditions. Compared to a fluid at rest, we foundthat in a shear flow, swimmers prefer the pull configuration of flagella over the wrapped one (bothemerging under CW flagellar rotation). Moreover, swimmers tend to align in parallel to the flow
direction, in contrast to a non-motile mutant (control with the same geometry), which does not showa preferred orientation under the same shear flow conditions. As the next step, to test an instantresponse of bacterial swimmers to flow, we developed biocompatible light-induced flow techniques
to locally advect or trap stained cells.3 Finally, with the help of AFM, we aim at specifying the nanomechanical properties of flagella and cell body to develop a mechanistic understanding of flagellar
dynamics under flow conditions in the future.
Our research demonstrates how bacteria adapt their swimming pattern to different
environmental conditions on the single-cell level. Understanding bacterial behavior and adaptation insuch conditions can be crucial for clarifying the processes of spreading and infection in capillaries aswell as biofilm formation in a natural environment.

!

Figure 1. (a) Schematic of bacterial swimming in a laminar shear flow. (b) Distribution of swimming modes under flow and no-flow
conditions

References:
1
Z. Alirezaeizanjani, R. Großmann, V. Pfeifer, M. Hintsche, and C. Beta, Sci. Adv. 6, (2020).
2
M. Hintsche, V. Waljor, R. Großmann, M.J. Kühn, K.M. Thormann, F. Peruani, and C. Beta, Sci.
Rep. 7, 1 (2017).
3
V. Muraveva, M. Bekir, N. Lomadze, R. Großmann, C. Beta, and S. Santer, Appl. Phys. Lett. 120,231905 (2022).

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Financial support
Travel and accommodation expenses will be supported up to 200K JPY fordomestic applicants / 350K JPY for overseas applicants in accordance with
Kanazawa University’s Travel Expense Regulations. Please contact us for details.
Å More info.
https://nanolsi.kanazawa-u.ac.jp/en/research/applications/bio-spm/

Visiting Fellows ProgramThis new program has been established for invited principal investigators andindependent researchers in molecular, cellular, or structural biology to gain experience
with HS-AFM, FM-AFM/3D-AFM, and SICM. This can also catalyze international
scientific cooperation between their institutions and NanoLSI.
Period of stayMore than one month (e.g., sabbatical leave)

Financial support
NanoLSI covers travel expenses, accommodation, and daily allowance inaccordance with the Kanazawa University’s Travel Regulations.
Applicants can be accompanied by up to two others such as fellow researchers,
postdocs from their own lab.
Å More info.
https://nanolsi.kanazawa-u.ac.jp/en/research/applications/visiting-fellows/58

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Organized by
Carsten Beta, Organizer University of Potsdam
Clemens Franz, Co-Organizer
Nano Life Science Institute (WPI-NanoLSI), Kanazawa University

Co-Organized by
Cancer Research Institute, Kanazawa University
Institute for Frontier Science Initiative, Kanazawa University

Contact
Secretariat
NanoLSI Administrative Officenanolsi-office@adm.kanazawa-u.ac.jp

Websitehttps:/nanolsi.kanazawa-u.ac.jp/en/7th-symposium/