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Video rate spectral fluorescence lifetime imaging with a 512 x 16 SPAD line sensor
Author(s): Ahmet T. Erdogan; Neil Finlayson; Gareth O. S. Williams; Elvira Williams; Robert K. Henderson
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Paper Abstract

We demonstrate a 512 x 16 CMOS single photon avalanche diode (SPAD) line sensor with per-pixel on-chip histogramming for video rate spectral fluorescence lifetime imaging (sFLIM). On-chip histogramming provides 32-bin histograms per pixel with 11bit/bin dynamic range. In addition, bin widths in time can be programmed from 51.20 ps to 6.55 ns, providing a histogram range from 1.64 ns to 209.72 ns to suit a wide range of fluorescence decays. At the end of a user defined exposure time, the full histogram data (i.e. 32-bins/pixel and 512 pixels) is first transferred to a FPGA in 84.48 μs via 64 data I/O pads at a 33.33 MHz I/O rate. The sensor data is then binned into two user defined spectral bands to provide spectral separation between different fluorophores, before being transferred to a PC via a USB3 connection for further processing. Fluorescence lifetimes for each spectral band are then rapidly estimated in software by applying the Centre-of-Mass Method (CMM), providing two 128 x 128 size spectral lifetime images in 1.384 s (i.e. with a frame rate of 0.72 fps). The frame rate can be increased by reducing the number of bins, reaching a maximum frame rate when only 2 bins are used with the Rapid Lifetime Determination (RLD) algorithm. In this paper we study the lifetime accuracy vs frame rate trade-offs by varying the number of histogram bins while carefully adjusting the bin widths for maximum bin counts. We validate the results using a Rhodamine 110 and Rhodamine B mixture solution which we separate them spectrally by their fluorescence lifetimes.

Paper Details

Date Published: 4 March 2019
PDF: 10 pages
Proc. SPIE 10889, High-Speed Biomedical Imaging and Spectroscopy IV, 108890M (4 March 2019); doi: 10.1117/12.2509607
Show Author Affiliations
Ahmet T. Erdogan, The Univ. of Edinburgh (United Kingdom)
Neil Finlayson, The Univ. of Edinburgh (United Kingdom)
Gareth O. S. Williams, The Univ. of Edinburgh (United Kingdom)
Elvira Williams, Durham Univ. (United Kingdom)
Robert K. Henderson, The Univ. of Edinburgh (United Kingdom)

Published in SPIE Proceedings Vol. 10889:
High-Speed Biomedical Imaging and Spectroscopy IV
Kevin K. Tsia; Keisuke Goda, Editor(s)

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