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

Ivan Michel Antolovic, Samuel Burri, Claudio Bruschini, Ron A. Hoebe, Edoardo Charbon

Research output: Contribution to journalArticleScientificpeer-review

21 Citations (Scopus)
52 Downloads (Pure)

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.

Original languageEnglish
Article number44108
Pages (from-to)1-11
Number of pages11
JournalScientific Reports
Volume7
DOIs
Publication statusPublished - 13 Mar 2017

Keywords

  • Electrical and electronic engineering
  • Imaging and sensing
  • Super-resolution microscopy

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