Standard Document
Second Edition
Hematology

CLSI H44

Methods for Reticulocyte Counting (Automated Blood Cell Counters, Flow Cytometry, and Supravital Dyes)

This document provides comprehensive protocols for reticulocyte counting using flow cytometry and automated hematology instruments. It includes detailed procedures for calibration, quality control, precision assessment, and the reference new methylene blue (NMB) method to ensure accurate and reliable results.

February 01, 2004
Charles F. Arkin, MD

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Abstract

Clinical and Laboratory Standards Institute document H44-A2—Methods for Reticulocyte Counting (Automated Blood Cell Counters, Flow Cytometry, and Supravital Dyes); Approved Guideline—Second Edition provides guidance for the performance of reticulocyte counting by flow cytometry and automated hematology instruments. This guideline addresses methods for determining the precision and trueness of the flow cytometer and blood cell counters based upon principles of focused flow dynamics, along with recommendations for calibration and quality control. A description of the new methylene blue (NMB) method, a method against which the test instrument can be compared, is also included. Additional topics discussed include reference intervals and use of related reticulocyte parameters, i.e., the immature reticulocyte fraction (formerly termed “reticulocyte maturation index”) and reticulocyte hemoglobin content (CHr).

Scope
This document outlines the essential factors that affect automated reticulocyte counting using flow cytometric principles. The designated Class C comparative method (see Section 4, Definitions) is the new methylene blue (NMB) staining of blood collected in ethylenediaminetetraacetic acid (EDTA) anticoagulant. Methods to ensure acceptable precision and trueness of the calibration and quality control of the automated reticulocyte counting methods are outlined. Procedures to develop appropriate reference ranges are also outlined. Methods used to express the relative and absolute maturation of reticulocytes are discussed, primarily in an effort to guide in the standardization of these additional clinically useful reticulocyte parameters.
Product Details
H44A2E
1-56238-527-5
39
Additional Details

This archived document is no longer being reviewed through the CLSI Consensus Document Development Process. However, this document is technically valid and because of its value to the laboratory community, it is being retained in CLSI’s library.

This document is available in electronic format only.

Authors
Charles F. Arkin, M.D., Chairholder
Frank M. LaDuca, Ph.D.
Bruce H. Davis, M.D., Vice-Chairholder
Ginette Y. Michaud, M.D.
J. David Bessman, M.D.
Onno W. van Assendelft, M.D., Ph.D.
Berend Houwen, M.D., Ph.D.
Abstract

Clinical and Laboratory Standards Institute document H44-A2—Methods for Reticulocyte Counting (Automated Blood Cell Counters, Flow Cytometry, and Supravital Dyes); Approved Guideline—Second Edition provides guidance for the performance of reticulocyte counting by flow cytometry and automated hematology instruments. This guideline addresses methods for determining the precision and trueness of the flow cytometer and blood cell counters based upon principles of focused flow dynamics, along with recommendations for calibration and quality control. A description of the new methylene blue (NMB) method, a method against which the test instrument can be compared, is also included. Additional topics discussed include reference intervals and use of related reticulocyte parameters, i.e., the immature reticulocyte fraction (formerly termed “reticulocyte maturation index”) and reticulocyte hemoglobin content (CHr).

Scope
This document outlines the essential factors that affect automated reticulocyte counting using flow cytometric principles. The designated Class C comparative method (see Section 4, Definitions) is the new methylene blue (NMB) staining of blood collected in ethylenediaminetetraacetic acid (EDTA) anticoagulant. Methods to ensure acceptable precision and trueness of the calibration and quality control of the automated reticulocyte counting methods are outlined. Procedures to develop appropriate reference ranges are also outlined. Methods used to express the relative and absolute maturation of reticulocytes are discussed, primarily in an effort to guide in the standardization of these additional clinically useful reticulocyte parameters.
Additional Details

This archived document is no longer being reviewed through the CLSI Consensus Document Development Process. However, this document is technically valid and because of its value to the laboratory community, it is being retained in CLSI’s library.

This document is available in electronic format only.

Authors
Charles F. Arkin, M.D., Chairholder
Frank M. LaDuca, Ph.D.
Bruce H. Davis, M.D., Vice-Chairholder
Ginette Y. Michaud, M.D.
J. David Bessman, M.D.
Onno W. van Assendelft, M.D., Ph.D.
Berend Houwen, M.D., Ph.D.