Publication

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

Edoardo Charbon, Claudio Bruschini, Samuel Burri, Ivan Michel Antolovic
2017
Journal Articles

Abstract

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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Edoardo Charbon, Claudio Bruschini, Samuel Burri, Ivan Michel Antolovic
2017
Journal Articles

Abstract

Official source

https://infoscience.epfl.ch/entities/publication/bd2d1e38-b8ea-4879-b5e8-9254f3e8e10d

About this result

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