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Flow Cytometry

Flow Cytometry

The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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Flow cytometry assay modifications: Recommendations for method validation based on CLSI H62 guidelines.

Sara A Monaghan¹, Steven Eck², Silvia Bunting³

¹Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, Pennsylvania, USA.

Cytometry. Part B, Clinical Cytometry

|August 21, 2024

Summary

This summary is machine-generated.

This study provides practical guidance on validating modified flow cytometry assays, building upon CLSI H62 recommendations. It details how assay modifications impact performance and suggests validation strategies for laboratory-initiated revisions.

Keywords:

CLSI H62 assay modification flow cytometry validation

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Area of Science:

Clinical Laboratory Science
Biotechnology
Analytical Chemistry

Background:

The Clinical and Laboratory Standards Institute (CLSI) H62 guideline (2021) offers recommendations for flow cytometry assay validation.
CLSI H62 addresses platform workflow, quality systems, instrument setup, assay development, and analytical method validation.
The guideline also includes recommendations for validating modified flow cytometric assays.

Purpose of the Study:

To expand on CLSI H62 recommendations for validating modified flow cytometry assays.
To discuss the impact of various assay modifications on overall assay performance.
To provide practical recommendations for laboratory-initiated assay revisions and validation strategies.

Main Methods:

Building upon existing CLSI H62 recommendations.
Analyzing the impact of different types of assay modifications on assay performance.
Proposing validation parameter evaluations based on the nature of the modification.
Discussing the influence of assay modification on the assay's intended use.

Main Results:

Different assay modifications have varying impacts on flow cytometry assay performance.
Specific validation parameters should be evaluated based on the type and extent of the assay modification.
Expert opinion provides rationale for minor deviations from CLSI H62 for laboratory-initiated revisions.

Conclusions:

This manuscript offers practical recommendations for validating modified flow cytometry assays, complementing CLSI H62.
The provided guidance aims to assist laboratories in navigating assay revisions while considering performance and intended use.
These recommendations are intended as practical guidance and not as a restrictive or universally sufficient standard for regulatory compliance.