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Calibration of flow cytometry detector systems

|November 1, 1981

Summary

This study introduces a calibration method for fluorescence activated cell sorters. It enables accurate measurement of relative fluorescence signal intensity across a wide dynamic range.

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Area of Science:

Biotechnology
Analytical Chemistry
Cell Biology

Background:

Accurate quantification of fluorescence signals is crucial for cell analysis.
Existing methods may struggle with wide dynamic ranges in fluorescence intensity.

Purpose of the Study:

To develop a robust calibration method for fluorescence activated cell sorters.
To enable precise determination of relative fluorescence signal intensity.

Main Methods:

A multistep calibration procedure was developed.
Evaluated signal gain from the amplifier module.
Determined photomultiplier response versus tube voltage for specific fluorochrome/filter combinations.

Main Results:

A simple mathematical calculation was derived from calibration data.
The method accurately determines relative fluorescence signal intensity.
Effective for samples with intensity differences spanning several orders of magnitude.

Conclusions:

The described method provides reliable relative fluorescence intensity measurements.
Enhances the quantitative capabilities of fluorescence activated cell sorting.
Applicable across diverse fluorochrome and filter configurations.

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