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  4. SPAD imagers for super resolution localization microscopy enable analysis of fast fluorophore blinking
 
research article

SPAD imagers for super resolution localization microscopy enable analysis of fast fluorophore blinking

Antolovic, Ivan Michel
•
Burri, Samuel  
•
Bruschini, Claudio  
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2017
Scientific Reports

sCMOS imagers are currently utilized (replacing EMCCD imagers) to increase the acquisition speed in super resolution localization microscopy. Single-photon avalanche diode (SPAD) imagers feature frame rates per bit depth comparable to or higher than sCMOS imagers, while generating microsecond 1-bit-frames without readout noise, thus paving the way to in-depth time-resolved image analysis. High timing resolution can also be exploited to explore fluorescent dye blinking and other photophysical properties, which can be used for dye optimization. We present the methodology for the blinking analysis of fluorescent dyes on experimental data. Furthermore, the recent use of microlenses has enabled a substantial increase of SPAD imager overall sensitivity (12-fold in our case), reaching satisfactory values for sensitivity-critical applications. This has allowed us to record the first super resolution localization microscopy results obtained with a SPAD imager, with a localization uncertainty of 20 nm and a resolution of 80 nm.

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Type
research article
DOI
10.1038/srep44108
Web of Science ID

WOS:000396422600001

Author(s)
Antolovic, Ivan Michel
Burri, Samuel  
Bruschini, Claudio  
Hoebe, Ron A.
Charbon, Edoardo  
Date Issued

2017

Publisher

Nature Research

Published in
Scientific Reports
Volume

7

Article Number

44108

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
AQUA  
Available on Infoscience
May 1, 2017
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/136834
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