Enumeration of Immunologically Defined Cell Populations by Flow Cytometry; Approved Guideline
|Publication Date:||1 May 2007|
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-b
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.