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  4. Analyzing blinking effects in super resolution localization microscopy with single-photon SPAD imagers
 
conference paper

Analyzing blinking effects in super resolution localization microscopy with single-photon SPAD imagers

Antolovic, Ivan Michel
•
Burri, Samuel  
•
Bruschini, Claudio  
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Enderlein, J
•
Gregor, I
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2016
Single Molecule Spectroscopy And Superresolution Imaging Ix
Single Molecule Spectroscopy and Superresolution Imaging IX

For many scientific applications, electron multiplying charge coupled devices (EMCCDs) have been the sensor of choice because of their high quantum efficiency and built-in electron amplification. Lately, many researchers introduced scientific complementary metal-oxide semiconductor (sCMOS) imagers in their instrumentation, so as to take advantage of faster readout and the absence of excess noise. Alternatively, single-photon avalanche diode (SPAD) imagers can provide even faster frame rates and zero readout noise. SwissSPAD is a 1-bit 512x128 SPAD imager, one of the largest of its kind, featuring a frame duration of 6.4 mu s. Additionally, a gating mechanism enables photosensitive windows as short as 5 ns with a skew better than 150 ps across the entire array. The SwissSPAD photon detection efficiency (PDE) uniformity is very high, thanks on one side to a photon-to-digital conversion and on the other to a reduced fraction of "hot pixels" or "screamers", which would pollute the image with noise. A low native fill factor was recovered to a large extent using a microlens array, leading to a maximum PDE increase of 12x. This enabled us to detect single fluorophores, as required by ground state depletion followed by individual molecule return imaging microscopy (GSDIM). We show the first super resolution results obtained with a SPAD imager, with an estimated localization uncertainty of 30 nm and resolution of 100 nm. The high time resolution of 6.4 mu s can be utilized to explore the dye's photophysics or for dye optimization. We also present the methodology for the blinking analysis on experimental data.

  • Details
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Type
conference paper
DOI
10.1117/12.2211430
Web of Science ID

WOS:000380592900004

Author(s)
Antolovic, Ivan Michel
Burri, Samuel  
Bruschini, Claudio  
Hoebe, Ron
Charbon, Edoardo  
Editors
Enderlein, J
•
Gregor, I
•
Gryczynski, Zk
•
Erdmann, R
•
Koberling, F
Date Issued

2016

Publisher

Spie-Int Soc Optical Engineering

Publisher place

Bellingham

Published in
Single Molecule Spectroscopy And Superresolution Imaging Ix
ISBN of the book

978-1-62841-948-1

Total of pages

7

Series title/Series vol.

Proceedings of SPIE

Volume

9714

Start page

971406

Subjects

fluorescence microscopy

•

image quality assessment

•

imaging ultrafast phenomena

•

photodetectors

•

photon-counting

•

super resolution

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
AQUA  
Event nameEvent placeEvent date
Single Molecule Spectroscopy and Superresolution Imaging IX

San Francisco, CA

FEB 13-14, 2016

Available on Infoscience
October 18, 2016
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/130091
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