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2017 ; 7
(ä): 44108
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SPAD imagers for super resolution localization microscopy enable analysis of fast
fluorophore blinking
#MMPMID28287122
Antolovic IM
; Burri S
; Bruschini C
; Hoebe RA
; Charbon E
Sci Rep
2017[Mar]; 7
(ä): 44108
PMID28287122
show ga
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.