Title
Direct frequency domain fluorescence lifetime imaging using field programmable gate arrays for real time processing
Date Issued
01 March 2020
Access level
open access
Resource Type
journal article
Author(s)
Serafino M.J.
Applegate B.E.
Texas A and M University
Publisher(s)
American Institute of Physics Inc.
Abstract
Frequency domain (FD) fluorescence lifetime imaging (FLIM) involves the excitation of the sample of interest with a modulated light source and digitization of the fluorescence emission for further analysis. Traditional FD-FLIM systems use heterodyne or homodyne detection, where the excitation light source and detector are modulated at specific frequency(s). More recently, FD-FLIM systems that use reflection of the light source as a trigger or phase reference for lifetime calculations have been developed. These detection schemes, however, require extra components that increase the cost and complexity of the FD-FLIM system. Here, we report a novel FD-FLIM detection scheme whereby the light source modulation and emission digitization are implemented using Field Programmable Gate Arrays (FPGAs), and fixed gain avalanche photodiodes are used for fluorescence detection. The reported FD-FLIM system was designed for probing nanosecond lifetime fluorophores (2-10 ns) at three emission bands simultaneously. The system utilizes a 375 nm diode laser for excitation at multiple simultaneous modulation frequencies (between 1 MHz and 83 MHz, bandwidth limited intentionally by using a lowpass filter) and three fixed gain avalanche photodiodes for simultaneous detection of three emission bands: 405/20 nm, 440/40 nm, and 525/50 nm (center/FWHM). Real-time computation of the modulation and phase lifetimes is simply performed by direct application of the discrete Fourier transform (max. of 10 frequencies) to the digitized fluorescence emission signals. The accuracy and sensitivity of this novel FD-FLIM detection scheme was demonstrated by imaging standard fluorophores and ex vivo unfixed human coronary artery tissue samples.
Volume
91
Issue
3
Number
033708
Language
English
OCDE Knowledge area
Ciencias de la información Robótica, Control automático
Scopus EID
2-s2.0-85082395450
PubMed ID
Source
Review of Scientific Instruments
ISSN of the container
00346748
Sponsor(s)
This work was funded by NIH (Grant Nos. 1R01CA218739 and 1R01HL111361). Postmortem human coronary artery segments were provided by Dr. Jessie Adame MD (Autopsy and Pathology Services, Houston, TX, USA). Lab space for imaging artery segments was provided by Dr. Brian Walton MD (Memorial Hermann Texas Medical Center, Houston, TX, USA). National Institutes of Health 1R01CA218739 NIH National Heart, Lung, and Blood Institute R01HL111361 NHLBI
Sources of information: Directorio de Producción Científica Scopus