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Detection of Protein Aggregation using Fluorescence Correlation Spectroscopy

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Published on: April 25, 2021

Small Particle Fluorescence and Light Scatter Calibration Using FCM_PASS Software.

Joshua A Welsh¹, Jennifer C Jones¹

¹Laboratory of Pathology, Translational Nanobiology Section, Center for Cancer Research, National Institute of Health, National Cancer Institute, Bethesda, Maryland.

Current Protocols in Cytometry

|September 16, 2020

Summary

This summary is machine-generated.

This study introduces FCM_PASS, a software for standardizing small particle flow cytometry. It enables reproducible analysis of extracellular vesicles (EVs) by calibrating fluorescence and light scatter data.

Keywords:

calibration extracellular vesicles fluorescence light scatter standardization

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Area of Science:

Biotechnology
Analytical Chemistry
Cell Biology

Background:

Flow cytometry is increasingly used for small particle analysis, including extracellular vesicles (EVs).
Lack of standardization in EV flow cytometry data reporting hinders interpretation, validation, and reproducibility.
Established methods exist for fluorescence and light scatter standardization.

Purpose of the Study:

To present FCM_PASS, a software package for standardizing small particle flow cytometry.
To enable fluorescence and light scatter calibration for small particles.
To generate reports conforming to the MIFlowCyt-EV standard.

Main Methods:

The study details a workflow for implementing calibration using FCM_PASS.
This includes acquiring and cataloging calibration materials, creating cytometer databases, and importing FCS files.
The process involves inputting fluorescence and light scatter calibration parameters and applying them to FCS files.

Main Results:

FCM_PASS provides a standardized approach to small particle flow cytometry calibration.
The software facilitates the generation of reports aligned with the MIFlowCyt-EV standard.
This promotes better data interpretation, validation, and reproducibility in EV research.

Conclusions:

FCM_PASS addresses the critical need for standardization in small particle flow cytometry.
Implementation of this software can significantly improve the quality and comparability of EV research data.
Standardized calibration is essential for advancing the field of extracellular vesicle analysis.