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Open Source High Content Analysis Utilizing Automated Fluorescence Lifetime Imaging Microscopy

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Published on: January 18, 2017

Instrument response standard in time-resolved fluorescence.

Rafal Luchowski¹, Zygmunt Gryczynski, Pabak Sarkar

¹Department of Molecular Biology and Immunology, Center for Commercialization of Fluorescence Technologies, UNTHSC, Fort Worth, Texas 76107, USA.

The Review of Scientific Instruments

|April 2, 2009

Summary

This summary is machine-generated.

The fluorescence lifetime of LDS 798 dye, measured at 24 picoseconds, offers a more accurate instrument response function approximation. This finding is crucial for precise fluorescence lifetime measurements, especially with near-infrared detectors.

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Area of Science:

Photophysics
Spectroscopy
Materials Science

Background:

Fluorescence lifetime measurements are critical in various scientific disciplines.
The instrument response function (IRF) is essential for accurate lifetime determination.
Conventional IRF approximation using scattered excitation light can introduce systematic errors due to wavelength-dependent detector response.

Purpose of the Study:

To investigate the suitability of LDS 798 dye fluorescence decay as an alternative IRF approximation.
To evaluate the accuracy of this method, particularly for near-infrared avalanche photodiodes.

Main Methods:

Measurement of the fluorescence lifetime of LDS 798 dye in aqueous solution.
Analysis of the time response characteristics of photon counting detectors.
Comparison of LDS 798 fluorescence decay with scattered excitation light for IRF approximation.

Main Results:

LDS 798 dye exhibits a short fluorescence lifetime of 24 ps, independent of excitation wavelength.
The fluorescence decay of LDS 798 provides a more accurate IRF approximation compared to scattered excitation light.
This improved approximation is particularly relevant for avalanche photodiodes in the near-infrared region.

Conclusions:

The fluorescence decay of LDS 798 dye is a superior method for approximating the IRF in fluorescence lifetime measurements.
This technique minimizes systematic errors associated with wavelength-dependent detector responses.
The findings enhance the precision of fluorescence lifetime studies, especially in the near-infrared spectral range.