APPENDIX 3C Wright-Giemsa and Nonspecific Esterase Staining of Cells
Published Online: 1 MAY 2001
Copyright © 2003 by John Wiley and Sons, Inc.
Lab Protocol Title
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.
- Published Online: 1 MAY 2001
- 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.