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  4. Phasor-based widefield FLIM using a gated 512×512 single-photon SPAD imager
 
conference paper

Phasor-based widefield FLIM using a gated 512×512 single-photon SPAD imager

Ulku, Arin Can
•
Bruschini, Claudio  
•
Antolovic, Ivan Michel  
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Periasamy, Ammasi
•
So, Peter T. C.
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February 22, 2019
Multiphoton Microscopy in the Biomedical Sciences XIX
Multiphoton Microscopy in the Biomedical Sciences XIX

Single-photon avalanche diode (SPAD) imagers can perform fast time-resolved imaging in a compact form factor, by exploiting the processing capability and speed of integrated CMOS electronics. Developments in SPAD imagers have recently made them compatible with widefield microscopy, thanks to array formats approaching one megapixel and sensitivity and noise levels approaching those of established technologies. In this paper, phasor-based FLIM is demonstrated with a gated binary 512×512 SPAD imager, which can operate with a gate length as short as 5.75 ns, a minimum gate step of 17.9 ps, and up to 98 kfps readout rate (1-bit frames). Lifetimes of ATTO 550 and Rhodamine 6G (R6G) solutions were measured across a 472×256 sub-array using phasor analysis, acquiring data by shifting a 13.1 ns gate window across the 50 ns laser period. The measurement accuracy obtained when employing such a scheme based on long, overlapping gates was validated by comparison with TCSPC measurements and fitting analysis results based on a standard Levenberg-Marquardt algorithm (<90% accuracy for the lifetime of R6G and ATTO 550). This demonstrates the ability of the proposed method to measure short lifetimes without minimum gate length requirements. The FLIM frame rate of the overall system can be increased up to a few fps for phasor-based widefield FLIM (moving closer to real-time operation) by FPGA-based parallel computation with continuous acquisition at the full speed of 98 kfps.

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Type
conference paper
DOI
10.1117/12.2511148
Author(s)
Ulku, Arin Can
Bruschini, Claudio  
Antolovic, Ivan Michel  
Weiss, Shimon
Michalet, Xavier
Charbon, Edoardo
Editors
Periasamy, Ammasi
•
So, Peter T. C.
•
König, Karsten
Date Issued

2019-02-22

Published in
Multiphoton Microscopy in the Biomedical Sciences XIX
Series title/Series vol.

Proceedings SPIE; 10882

Start page

21

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
AQUA  
Event nameEvent placeEvent date
Multiphoton Microscopy in the Biomedical Sciences XIX

San Francisco, CA, USA

2-7 February, 2019

Available on Infoscience
August 2, 2019
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/159522
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