CLSI H42
Enumeration of Immunologically Defined Cell Populations by Flow Cytometry
This guideline outlines essential procedures and quality assurance measures for immunophenotypic analysis using flow cytometry. It standardizes practices across laboratories by covering specimen handling, instrument calibration, staining protocols, and data analysis to ensure accurate and comparable results.
This reaffirmed document has been reviewed and confirmed as suitable to remain published without revision to content, as ofJune 2017.
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{{FormatPrice(nonMemberPrice)}} List PriceClinical and Laboratory Standards Institute document H42-A2—Enumeration of Immunologically Defined Cell Populations by Flow Cytometry; Approved Guideline—Second Edition was developed to address issues of procedures and quality assurance for clinical applications of flow cytometry. It is designed to aid clinical laboratorians in the development of quality assurance procedures and to establish the foundation for different laboratories using different commercially available instruments to obtain comparable results. Specific topics covered include: specimen collection, transport, and preparation; sample quality control and staining procedures; instrument calibration; sample analysis; and data analysis, storage, and reporting.
The scope of this document is to establish performance guidelines for the identification and enumeration of lymphocyte subpopulations and the enumeration of CD34+ hematopoietic progenitors using immunofluorescence-based flow cytometry (FCM).
The working group recognizes that other, so-called nontraditional methodologies exist or are in development for enumeration of CD4+ T-lymphocytes (e.g., systems using microcapillary sample delivery or nonfluorescent cell detection; see Appendix C). Some of the issues discussed in this document are common to the use of any method for CD4+ T-cell enumeration (e.g., sample collection and transport, safety issues, data reporting, and interpretation). However, issues such as sample preparation, instrument calibration, and quality control differ significantly for nontraditional methodologies and are not discussed in this document.
Presently, there are no universally accepted standards for precision, accuracy, and interlaboratory comparability in lymphocyte enumeration by FCM. General consensus was reached on the basic International Society for Hematotherapy and Graft Engineering (ISHAGE)9 guidelines for CD34 analysis, and this forms the basis of the technique described herein. It is beyond the scope of this document to establish general performance criteria and reference intervals. Therefore, it is each laboratory’s responsibility to establish instrument performance criteria and staining characteristics for its own specific reagents.
This document is available in electronic format only.
The U.S. Food and Drug Administration (FDA) has evaluated and recognized this approved-level consensus standard for use in satisfying a regulatory requirement.
Clinical and Laboratory Standards Institute document H42-A2—Enumeration of Immunologically Defined Cell Populations by Flow Cytometry; Approved Guideline—Second Edition was developed to address issues of procedures and quality assurance for clinical applications of flow cytometry. It is designed to aid clinical laboratorians in the development of quality assurance procedures and to establish the foundation for different laboratories using different commercially available instruments to obtain comparable results. Specific topics covered include: specimen collection, transport, and preparation; sample quality control and staining procedures; instrument calibration; sample analysis; and data analysis, storage, and reporting.
The scope of this document is to establish performance guidelines for the identification and enumeration of lymphocyte subpopulations and the enumeration of CD34+ hematopoietic progenitors using immunofluorescence-based flow cytometry (FCM).
The working group recognizes that other, so-called nontraditional methodologies exist or are in development for enumeration of CD4+ T-lymphocytes (e.g., systems using microcapillary sample delivery or nonfluorescent cell detection; see Appendix C). Some of the issues discussed in this document are common to the use of any method for CD4+ T-cell enumeration (e.g., sample collection and transport, safety issues, data reporting, and interpretation). However, issues such as sample preparation, instrument calibration, and quality control differ significantly for nontraditional methodologies and are not discussed in this document.
Presently, there are no universally accepted standards for precision, accuracy, and interlaboratory comparability in lymphocyte enumeration by FCM. General consensus was reached on the basic International Society for Hematotherapy and Graft Engineering (ISHAGE)9 guidelines for CD34 analysis, and this forms the basis of the technique described herein. It is beyond the scope of this document to establish general performance criteria and reference intervals. Therefore, it is each laboratory’s responsibility to establish instrument performance criteria and staining characteristics for its own specific reagents.
This document is available in electronic format only.
The U.S. Food and Drug Administration (FDA) has evaluated and recognized this approved-level consensus standard for use in satisfying a regulatory requirement.