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Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy

Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy

Published on: January 18, 2017

Fluorescence Lifetime Measurements and Analyses: Protocols Using Flow Cytometry and High-Throughput Microscopy.

Jessica P Houston¹, Samantha Valentino², Aric Bitton³

¹Department of Chemical & Materials Engineering, New Mexico State University, Las Cruces, NM, USA. jph@nmsu.edu.

Methods in Molecular Biology (Clifton, N.J.)

|March 25, 2024

Summary

Fluorescence lifetime measurements offer a powerful, quantitative parameter for cell analysis. This technique enhances cell counting, sorting, and imaging through advanced time-resolved flow cytometry and machine learning applications.

Keywords:

FRET Fluorescence lifetime Frequency domain Time domain Time-resolved flow cytometry

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Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells

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Published on: February 9, 2012

Area of Science:

Biophysics
Cell Biology
Analytical Chemistry

Background:

Fluorescence decay timing is a well-established parameter in cell analysis.
Traditional cytometry methods benefit from incorporating fluorescence lifetime measurements.
Understanding fluorescence decay kinetics is crucial for advanced cytometric applications.

Purpose of the Study:

To provide an overview of fluorescence lifetime measurement techniques in cytometry.
To explore how fluorescence lifetime enhances existing cytometry protocols.
To review modern high-throughput lifetime analysis methods.

Main Methods:

Review of theoretical background and history of time-resolved flow cytometry.
Analysis of applications benefiting from fluorescence lifetime as a photophysical trait.
Discussion of modern high-throughput scanning techniques including high-speed microscopy and machine learning.

Main Results:

Fluorescence lifetime provides quantitative, informative data for cell analysis.
Time-resolved flow cytometry has evolved significantly for enhanced cellular insights.
Modern techniques enable high-throughput fluorescence lifetime scanning.

Conclusions:

Fluorescence lifetime measurement is a valuable cytometric parameter.
Advanced techniques like machine learning are revolutionizing lifetime analysis.
This approach offers significant potential for future cell biology research.