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OptnanoATcryo SIGNED

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

Total Cost €


EC-Contrib. €






 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.

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

Project "OptnanoATcryo" data sheet

The following table provides information about the project.


Organization address
address: STEVINWEG 1
city: DELFT
postcode: 2628 CN

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


Take a look of project's partnership.

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


 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.


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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