Benzidine blood stain

Benzidine is a reagent that forms a blue precipitate upon oxidation by the heme group of hemoglobin in the presence of hydrogen peroxide. Thus it serves as a histochemical stain specific for differentiated red blood cells.

The figures are from A. Hemmatti-Brivanlou and G.H. Thomsen (1995). Ventral mesodermal patterning in Xenopus embryos: expression patterns and actvities of BMP-2 and BMP-4. Devl. Genet. 17; 78-89

This page contains previously published text and images. Copyright © to all such materials belongs to Wiley-Liss Inc. (1995).
Click on images to view enlargements (6K to 7K, < 2 seconds).

A.
Wild type embryo stained at stage 35. The ventral blood island (VBI) is demarcated by the presence of blue, stained erythrocytes.	A stage 50 tadpole stained with benzidine to show circulating erythrocytes in the major blood vessels and heart (H).	A control (pGEM-7 RNA-injected) albino animal cap stains negative with benzidine.	Animal cap injected with 2 ng of BMP-4 stains positive for erythrocytes (this is probably a fused pair of caps)

The staining protocol is:

Fix embryos or tissues for 5 minutes in 12% glacial acetic acid containing 0.4% benzidine (Sigma # B-3503 CARCINOGEN). Start reaction by addition of hydrogen peroxide to a final concentration of 0.3%. Incubate at RT (22 o C). Color should develop within 10-15 minutes. Alternatively, the specimens can be incubated in an acetic acid/benzidine solution already containing freshly added hydrogen peroxide. Monitor reaction for color development and photograph immediately. The blue precipitate is unstable in the presence of peroxide, and the peroxide will also gradually destroy the specimens. Peroxide can be washed out with 12% acetic acid and fixed in methanol to stablize the precipitate, but the color will turn brown and lose intensity. The protocol is adapted from Orkin S.H., Harosi, F.L. and Leder, P. (1975): Differentiation of erythroleukemic cells and their somatic hybrids. PNAS 72:98-102.

Cloned by:			Original description:
			Hemmati-Brivanlou A, Thomsen GH Dev Genet 1995;17(1):78-89
Images submitted by:			Sequence
Jerry Thomsen			N/A
Probe available from:			Other papers by:
N/A			A Hemmati-Brivanlou or GH Thomsen
In situ probe details:			Other papers on:
plasmid name:	antisense RE cut:	antisense transcription:
mRNA generation
plasmid name:	sense RE cut:	sense transcription:

community	gene expression	genomics	genetics	atlas/movies/fate	cell biology
literature	methods	events	magazine
indexes	genes by name	genes by site	antibodies by name	Abs by site	search

contact the site administrator

credit/copright information information