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Wide-field time-gated SPAD imager for phasor-based FLIM applications.

Arin Ulku1, Andrei Ardelean1, Michel Antolovic1

  • 1AQUA Lab, Ecole Polytechnique Fédérale de Lausanne (EPFL), Neuchâtel, Switzerland.

Methods and Applications in Fluorescence
|January 23, 2020
PubMed
Summary
This summary is machine-generated.

A new wide area time-gated single-photon avalanche diode (SPAD) array enables video-rate fluorescence lifetime imaging microscopy (FLIM). This detector performs as an ideal shot-noise-limited sensor, even with limited photons.

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Area of Science:

  • Biophotonics
  • Optical Imaging
  • Fluorescence Spectroscopy

Background:

  • Fluorescence Lifetime Imaging Microscopy (FLIM) is a powerful technique for analyzing biological samples.
  • High-throughput FLIM requires sensitive and fast detectors.
  • Single-Photon Avalanche Diode (SPAD) arrays offer potential for advanced FLIM applications.

Purpose of the Study:

  • To evaluate the performance of a novel wide-area time-gated SPAD array for phasor-FLIM.
  • To investigate the impact of gate length, gate number, and signal intensity on lifetime measurements.
  • To assess the suitability of the phasor approach for large-format SPAD sensors.

Main Methods:

  • Characterization of a 512x512 pixel time-gated SPAD array.
  • Phasor analysis applied to FLIM data acquired with varying experimental parameters.
  • Assessment of lifetime accuracy and precision under different signal conditions.

Main Results:

  • The SPAD array demonstrated performance close to the ideal shot-noise limit.
  • Accurate and precise fluorescence lifetime measurements were achieved.
  • Video-rate FLIM acquisition is feasible with this detector technology.
  • The phasor approach effectively managed large datasets from the 512x512 sensor.

Conclusions:

  • The wide-area time-gated SPAD array is a capable sensor for high-speed FLIM.
  • Phasor analysis is well-suited for processing data from large SPAD arrays, even with limited photons.
  • This technology advances the potential for rapid, quantitative biological imaging.