Time-zoomable FRET spectroscopy with a 512 x16 SPAD line sensor

Ahmet T. Erdogan; Gareth O. S. Williams; Andrea Usai; Nikola Krstajic; Neil Finlayson; Andrew Beavil; Rebecca Beavil; Robert K. Henderson

doi:10.1117/12.2290058

20 February 2018 Time-zoomable FRET spectroscopy with a 512 x16 SPAD line sensor

Ahmet T. Erdogan, Gareth O. S. Williams, Andrea Usai, Nikola Krstajic, Neil Finlayson, Andrew Beavil, Rebecca Beavil, Robert K. Henderson

Author Affiliations +

Proceedings Volume 10504, Biophysics, Biology and Biophotonics III: the Crossroads; 105040M (2018) https://doi.org/10.1117/12.2290058
Event: SPIE BiOS, 2018, San Francisco, California, United States

Abstract

We demonstrate a new 512x16 single photon avalanche diode (SPAD) based line sensor with per-pixel TCSPC histogramming for time-resolved, time-zoomable, FRET spectroscopy. The line sensor can operate in single photon counting (SPC) mode as well as time-correlated single photon counting (TCSPC) and per-pixel histogramming modes. TCSPC has been the preferred method for fluorescence lifetime measurements due to its collection of full decays as a histogram of arrival times. However, TCSPC is slow due to only capturing one photon per exposure and large timestamp data transfer requirements for offline histogramming. On-chip histogramming improves the data rate by allowing multiple SPAD pulses (up to one pulse per laser period) to be processed in each exposure cycle, along with secondly reducing the I/O bottleneck as only the final histogram is transferred. This can enable 50x higher acquisition rates (up to 10 billion counts per second), along with time-zoomable histogramming operation from 1.6ns to 205ns with 50ps resolution. A broad spectral range can be interrogated with the sensor (450-900nm). Overall, these sensors provide a unique combination of light sensing capabilities for use in high speed, sensitive, optical instrumentation in the time/wavelength domain. We test the sensor performance by observation of fluorescence resonance energy transfer (FRET) between FAM and TAMRA and between EGFP and RFP FRET standards.

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Ahmet T. Erdogan, Gareth O. S. Williams, Andrea Usai, Nikola Krstajic, Neil Finlayson, Andrew Beavil, Rebecca Beavil, and Robert K. Henderson "Time-zoomable FRET spectroscopy with a 512 x16 SPAD line sensor ", Proc. SPIE 10504, Biophysics, Biology and Biophotonics III: the Crossroads, 105040M (20 February 2018); https://doi.org/10.1117/12.2290058

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