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An Innovative Method for Exosome Quantification and Size Measurement

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Published on: January 17, 2015

Pulse width for particle sizing.

Robert A Hoffman¹

¹BD Biosciences, San Jose, California, USA.

Current Protocols in Cytometry

|October 10, 2009

Summary

This summary is machine-generated.

Optical pulse width in flow cytometry offers a reliable particle size measurement, independent of light scatter variations. This method enables accurate particle sizing through pulse width calibration, enhancing flow cytometry analysis.

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Simultaneous Measurement of Turbulence and Particle Kinematics Using Flow Imaging Techniques

Simultaneous Measurement of Turbulence and Particle Kinematics Using Flow Imaging Techniques

Published on: March 12, 2019

Area of Science:

Biophysics
Analytical Chemistry
Optical Physics

Background:

Flow cytometry commonly uses light scatter to estimate particle size, but this is influenced by factors like particle shape and refractive index.
Optical pulse width in flow cytometry offers an alternative particle size metric, potentially overcoming limitations of light scatter.
The relationship between pulse width and particle size is theoretically predictable, especially for fluorescence signals.

Purpose of the Study:

To establish protocols for amplitude-independent optical pulse width measurements in flow cytometry.
To develop methods for calibrating pulse width measurements to determine particle diameter.
To demonstrate the utility of pulse width as a quantitative particle size parameter.

Main Methods:

Determining the signal amplitude range for reliable, amplitude-independent pulse width measurements.
Implementing theoretical predictions for pulse width based on particle shape and fluorescence signals.
Calibrating pulse width measurements against known particle sizes to establish a quantitative scale.

Main Results:

Demonstrated that optical pulse width provides particle size information independent of common light scatter interferences.
Established protocols for amplitude-independent pulse width measurements across various signal ranges.
Showcased successful quantitative calibration of pulse width to particle diameter using fluorescence signals.

Conclusions:

Optical pulse width is a robust and independent parameter for particle sizing in flow cytometry.
The developed protocols and calibration methods enable accurate and reliable particle diameter determination.
This approach enhances the quantitative capabilities of flow cytometry for particle analysis.