APPENDIX 3C Wright-Giemsa and Nonspecific Esterase Staining of Cells

  1. Warren Strober

Published Online: 1 MAY 2001

DOI: 10.1002/0471142735.ima03cs21

Current Protocols in Immunology

Current Protocols in Immunology

How to Cite

Strober, W. 2001. Wright-Giemsa and Nonspecific Esterase Staining of Cells. Current Protocols in Immunology. 21:3C:A.3C.1–A.3C.3.

Author Information

  1. National Institute of Allergy and Infectious Diseases, Bethesda, Maryland

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: MAR 1997


This appendix provides two protocols for staining cells. The stains used in the Wright-Giemsa protocol, the Romanowsky stains, are a mixture of methylene blue (and other closely related thiazine dyes) and eosin. The staining protocol is a two-stage method that allows for a more intense staining of the nuclei than would be possible if the Wright and Giemsa stains were mixed together. In the second protocol, nonspecific esterase stain are used to identify cell types containing esterases that have a characteristic ability to split esters under particular conditions. In the staining method given here, the substrate, a-naphthyl butyrate, is incubated with cells under conditions in which esterases present in monocytes/macrophages split the substrate to yield an intermediate that can be coupled with a substance, hexazotized pararosaniline, to yield a colored precipitate. Thus, this staining reaction can be used to identify monocytes/macrophages in cell preparations.