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InCell SIGNED

High speed AFM imaging of molecular processes inside living cells

Total Cost €

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EC-Contrib. €

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Partnership

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 InCell project word cloud

Explore the words cloud of the InCell project. It provides you a very rough idea of what is the project "InCell" about.

viscous    pt    incell    closest    exists    functional    lab    afm    limited    revolutionize    microscopy    barrel    extraction    infection    ort    molecular    single    inside    leak    first    nuclear    coat    area    microfabricated    membrane    minimally    seals    tapping    cells    optical    changer    questions    generation    cantilever    imaging    parasitic    patch    game    outside    subtracted    fluid    technique    possibilities    direct    cytosol    photo    causing    remedy    resolution    endocytosis    tip    kinetics    plasma    interactions    assembly    enter    nanoscale    dream    shell    cell    unfortunately    thermal    speed    networks    encased    mode    signalling    biology    nanometre    tool    time    pits    combined    fundamental    vesicular    deflection    native    pore    clathrin    lattice    interaction    guide    true    viral    stability    signal    hs    fluids    resonance    onto    double    force    structural    observation    clamp    environment    transport    invasive    confocal    atomic    living    bacterial    dynamics    coated    bio    off    myriad    answer   

Project "InCell" data sheet

The following table provides information about the project.

Coordinator
ECOLE POLYTECHNIQUE FEDERALE DE LAUSANNE 

Organization address
address: BATIMENT CE 3316 STATION 1
city: LAUSANNE
postcode: 1015
website: www.epfl.ch

contact info
title: n.a.
name: n.a.
surname: n.a.
function: n.a.
email: n.a.
telephone: n.a.
fax: n.a.

 Coordinator Country Switzerland [CH]
 Total cost 1˙999˙925 €
 EC max contribution 1˙999˙925 € (100%)
 Programme 1. H2020-EU.1.1. (EXCELLENT SCIENCE - European Research Council (ERC))
 Code Call ERC-2017-COG
 Funding Scheme ERC-COG
 Starting year 2018
 Duration (year-month-day) from 2018-04-01   to  2023-03-31

 Partnership

Take a look of project's partnership.

# participants  country  role  EC contrib. [€] 
1    ECOLE POLYTECHNIQUE FEDERALE DE LAUSANNE CH (LAUSANNE) coordinator 1˙999˙925.00

Map

 Project objective

Imaging the inside of living cells with single nanometre resolution has been a long-standing dream in bio-microscopy. Direct observation of changes to molecular networks inside of living cells would revolutionize the way we study structural cell biology. Unfortunately, no such tool exists. Atomic force microscopy (AFM) is the closest we have, to nanoscale functional imaging of cells in their native, fluid environment. However, it is limited to imaging the outside of the cell. With InCell, I will remedy this by developing an AFM capable of imaging the inside of living cells. The approach is based on a microfabricated high speed AFM cantilever encased in a double barrel patch-clamp shell. The patch clamp shell seals onto the plasma membrane of the cell, so that the tip of the AFM cantilever can enter the cell without causing the cytosol to leak out. Parasitic interactions of the AFM tip with the cytosol will be subtracted from the cantilever deflection signal, using high speed photo-thermal off-resonance tapping (PT-ORT), a novel AFM mode we have recently developed in my lab. This allows the extraction of the true tip-sample interaction, even in viscous fluids. A dedicated InCell HS-AFM combined with confocal optical microscopy will be used to guide the InCell cantilever inside the cell to the area of interest. Using this minimally invasive technique we will study the formation of clathrin coated pits, a crucial part of endocytosis. By imaging for the first time the nanoscale dynamics of this process in living cells, we aim to answer fundamental questions about the clathrin coat assembly. We will characterize the kinetics, stability and force generation by the clathrin lattice. This will be the first example of how enabling nanoscale imaging inside living cells will be a game changer in cell biology. It will open up a myriad of possibilities for the study of vesicular transport, viral and bacterial infection, nuclear pore transport, cell signalling and many more.

 Publications

year authors and title journal last update
List of publications.
2019 Edward S. Parsons, George J. Stanley, Alice L. B. Pyne, Adrian W. Hodel, Adrian P. Nievergelt, Anaïs Menny, Alexander R. Yon, Ashlea Rowley, Ralf P. Richter, Georg E. Fantner, Doryen Bubeck, Bart W. Hoogenboom
Single-molecule kinetics of pore assembly by the membrane attack complex
published pages: , ISSN: 2041-1723, DOI: 10.1038/s41467-019-10058-7
Nature Communications 10/1 2019-12-16
2019 Adrian P. Nievergelt, Christoph Kammer, Charlène Brillard, Eva Kurisinkal, Maartje M. C. Bastings, Alireza Karimi, Georg E. Fantner
Large‐Range HS‐AFM Imaging of DNA Self‐Assembly through In Situ Data‐Driven Control
published pages: 1900031, ISSN: 2366-9608, DOI: 10.1002/smtd.201900031
Small Methods 2019-12-16
2019 Ke Liu, Chao Pan, Alexandre Kuhn, Adrian Pascal Nievergelt, Georg E. Fantner, Olgica Milenkovic, Aleksandra Radenovic
Detecting topological variations of DNA at single-molecule level
published pages: , ISSN: 2041-1723, DOI: 10.1038/s41467-018-07924-1
Nature Communications 10/1 2019-05-27
2018 Adrian Nievergelt, Charlène Brillard, Haig Eskandarian, John McKinney, Georg Fantner
Photothermal Off-Resonance Tapping for Rapid and Gentle Atomic Force Imaging of Live Cells
published pages: 2984, ISSN: 1422-0067, DOI: 10.3390/ijms19102984
International Journal of Molecular Sciences 19/10 2019-04-03
2018 Adrian P. Nievergelt, Niccolò Banterle, Santiago H. Andany, Pierre Gönczy, Georg E. Fantner
High-speed photothermal off-resonance atomic force microscopy reveals assembly routes of centriolar scaffold protein SAS-6
published pages: 696-701, ISSN: 1748-3387, DOI: 10.1038/s41565-018-0149-4
Nature Nanotechnology 13/8 2019-03-12

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The information about "INCELL" are provided by the European Opendata Portal: CORDIS opendata.

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