King's College London
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SPADlinearization.tgz (470.77 MB)

SPAD linearization code for processing calibration and measurement data from the SPAD camera

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posted on 2024-03-06, 16:46 authored by Jakub NedbalJakub Nedbal

The SPADlinearization software contains a Mathworks MATLAB script SPADcorrection.m. This is used to create calibration data from measurements of dark count map (dark sensor), instrument response function (pulsed light illuminated sensor), and code density map (constant light illuminated sensor) acquired on the microscope with the QuantiCam camera. The script produces calibration data. There are two calibration datasets in the file

  • binCorrection78MHz_NKT_chip12_Allura.full.mat
  • binCorrection78MHz_NKT_chip12_FITC.full.mat

Differing in the calibration solution (Allura Red or quenched fluorescein) and the fluorescence wavelength measured (667 nm and 535 nm, respectively). These calibration measurements are only suitable for the single sensor and optical setup used in the published experiments and is not generally applicable. The calibrations need to be done for each optical setup and sensor separately.

processFLIM.m is a script used to process fluroescence decay measurement data from the QuantiCam SPAD camera. This script was used to process all measurement data presented in the publication.

QuantiCAM_calibration_and_measurement_guide.pdf is a protocol explaining how calibration data is acquired and processed and how fluorescence lifetime measurements are processed.

Most recent version of the source code is available at:

https://www.github.com/jnedbal/SPADcorrection/

This software is a supplement to the article "A time-correlated single photon counting SPAD array camera with a bespoke data-processing algorithm for lightsheet fluorescence lifetime imaging (FLIM) and FLIM videos"

Funding

Development of wide-field TCSPC fluorescence microscopy for cell membrane studies

Biotechnology and Biological Sciences Research Council

Find out more...

EP/X525571/1

History

Data collection from date

4. May 2020

Data collection to date

7. Oct 2022

Copyright owner

Jakub Nedbal