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Updated: Apr 20, 2026

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 plate reader: resolution and precision meet high-throughput.

Karl J Petersen¹, Kurt C Peterson², Joseph M Muretta¹

¹Department of Biochemistry, Molecular Biology and Biophysics, University of Minnesota, Minneapolis, Minnesota 55455, USA.

The Review of Scientific Instruments

|November 29, 2014

Summary

This study presents a fast, high-precision nanosecond time-resolved fluorescence spectrometer for 384-well plates. The instrument enables rapid, accurate fluorescence lifetime measurements for high-throughput screening applications.

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Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy

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Published on: April 7, 2023

Area of Science:

Analytical Chemistry
Spectroscopy
Biophysics

Background:

Fluorescence lifetime measurements offer valuable insights into molecular environments.
Traditional methods can be time-consuming, limiting high-throughput applications.
Developing rapid, precise fluorescence lifetime instrumentation is crucial for screening.

Purpose of the Study:

To develop a nanosecond time-resolved fluorescence spectrometer for rapid, high-precision measurements.
To enable high-throughput fluorescence lifetime screening using a 384-well microplate format.
To achieve superior precision compared to simultaneous intensity measurements.

Main Methods:

Utilized high-energy pulsed laser sources (5-10 kHz) and a photomultiplier with a 1 GHz digitizer.
Employed direct waveform recording to capture fluorescence decay waveforms from each well.
Achieved an effective sample rate of 5 GHz for precise lifetime determination.

Main Results:

Acquired fluorescence decay waveforms from a 384-well plate in 3 minutes with signal-to-noise > 400.
Obtained fluorescence lifetime measurements 5- to 25-fold more precise than intensity measurements.
Resolved dye mixtures with better than 1% accuracy and measured lifetimes as short as 0.04 ns.

Conclusions:

The developed fluorescence lifetime plate reader facilitates rapid, multiple-well measurements.
Offers an acquisition time of 0.5 seconds per well, ideal for high-throughput screening.
Demonstrates the potential for advanced applications in drug discovery and biochemical assays.