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OptnanoATcryo SIGNED

Optical nanoscopy at 1 nm resolution: far-field fluorescence control at cryogenic temperatures

Total Cost €

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EC-Contrib. €

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Partnership

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 OptnanoATcryo project word cloud

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

big    identical    density    negligible    molecular    structures    fluorescence    relaxing    undemanding    nm    notably    function    samples    subcellular    offers    polarization    image    temperatures    schemes    perspective    localization    leap    data    technique    chromosomal    subsequent    ensures    promises    photobleaching    cryo    fewer    averaging    investigation    collected    resolutions    labels    fluorescent    introduce    twofold    depletion    nanoscopy    10    larger    revolution    microscopy    triplet    outcome    subnanometer    nanometer    dna    entities    prior    machinery    biology    electron    gene    photons    mean    structure    describe    biophysical    combination    realize    3d    priors    emitters    priori    cryogenic    isotropic    regulation    setup    labelling    ranging    imaging    stimulated    maintaining    counts    250    cell    experimental    sensitive    imaged    relatively    excitation    observations    resolution    orientational    super    conformation    conventional    structural    powerful    spectacular    emitter    optical    unprecedented    blinking    functional    reconstruction    shelving    packaging    photon    single    sparsity    employing    detection    models   

Project "OptnanoATcryo" data sheet

The following table provides information about the project.

Coordinator
TECHNISCHE UNIVERSITEIT DELFT 

Organization address
address: STEVINWEG 1
city: DELFT
postcode: 2628 CN
website: www.tudelft.nl

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 Netherlands [NL]
 Total cost 1˙911˙792 €
 EC max contribution 1˙911˙792 € (100%)
 Programme 1. H2020-EU.1.1. (EXCELLENT SCIENCE - European Research Council (ERC))
 Code Call ERC-2014-CoG
 Funding Scheme ERC-COG
 Starting year 2015
 Duration (year-month-day) from 2015-07-01   to  2020-06-30

 Partnership

Take a look of project's partnership.

# participants  country  role  EC contrib. [€] 
1    TECHNISCHE UNIVERSITEIT DELFT NL (DELFT) coordinator 1˙911˙792.00

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 Project objective

Optical nanoscopy is a powerful technique used in biology to study subcellular structures and function via specifically targeted fluorescent labels. Localization microscopy in particular offers a much better resolution (~10-50 nm) than conventional microscopy (~250 nm) while being relatively undemanding on the experimental setup and the subsequent image analysis. The next revolution in imaging to 1 nm isotropic resolution in 3D must realize a big increase in the number of collected photons from single fluorescent emitters as well as in the labelling density. Only then can subcellular structures be imaged at the molecular level to study the molecular machinery of the cell. Notably observations of DNA conformation in 3D at such resolutions would be spectacular and enable investigation of biophysical models ranging from chromosomal DNA packaging to gene regulation.

I propose a new imaging technique based on fluorescence control at cryogenic temperatures in combination with novel data driven super-resolution reconstruction schemes employing prior knowledge that promises this unprecedented optical far-field resolution. I introduce a twofold technical leap by i) much higher photon counts due to negligible photobleaching at cryogenic temperatures while maintaining the sparsity required for single emitter localization and ii) relaxing the required labelling density using a priori information and the averaging of many identical entities. Orientational blinking ensures single emitter localization via a combination of polarization sensitive excitation, detection and stimulated depletion and triplet state shelving. Biophysical models of cell structures and data driven priors mean that fewer samples are needed to fully describe a structure. In a larger perspective, the outcome of this research will enable the combination of structural cryo-electron microscopy imaging at subnanometer resolutions with functional fluorescent imaging at the nanometer scale.

 Publications

year authors and title journal last update
List of publications.
2019 Hamidreza Heydarian, Adrian Przybylski, Florian Schueder, Ralf Jungmann, Ben van Werkhoven, Jan Keller-Findeisen, Jonas Ries, Sjoerd Stallinga, Mark Bates, Bernd Rieger
Three dimensional particle averaging for structural imaging of macromolecular complexes by localization microscopy
published pages: , ISSN: , DOI: 10.1101/837575
bioarchive 2020-02-04
2017 B. Rieger and S. Stallinga
Data fusion at the nanoscale: Imaging at resolutions better than wavelength/100
published pages: 13-14, ISSN: 0926-4981, DOI:
ERCIM News: Special theme: Computational Imaging 2019-06-06
2018 C. Hulleman, M. Huisman, R. Moerland, D. Grünwald, S. Stallinga, B.Rieger
Fluorescence polarization control for on-off switching of single molecules at cryogenic temperatures
published pages: , ISSN: 2366-9608, DOI:
Small Methods 2019-06-06
2016 R. Heintzmann, P. Relich, R.P.J. Nieuwenhuizen, K.A. Lidke and B. Rieger
Calibrating photon counts from a single image
published pages: , ISSN: , DOI:
ArXiv 2019-06-06
2018 K. Martens, A.N. Bader, S. Baas, B. Rieger, J. Hohlbein
Phasor based single-molecule localization microscopy in 3D (pSMLM-3D): an algorithm for MHz localization rates using standard CPUs
published pages: 123311, ISSN: 0021-9606, DOI:
Journal of Chemical Physics 148 2019-06-06

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