[Histonet] Hemoglobin stain

[Martin, Erin]

Hi all,



I have a pathologist that is looking for a special stain that "highlights blood."  Initially she had wanted to use benzidine as referenced in the article below but our safety office let me know that we would need some pretty hefty air scrubbers and respirators to use it safely.  Does anyone have a protocol that might give the same results?



Thank you,

Erin Martin

Erin Martin, Histology Supervisor
UCSF  Dermatopathology Service
415-353-7248

Am J Dermatopathol. 1995 Aug;17(4):362-7.
Benzidine stain for the histochemical detection of hemoglobin in splinter
hemorrhage (subungual hematoma) and black heel.
Hafner J(1), Haenseler E, Ossent P, Burg G, Panizzon RG.
Author information:
(1)Department of Dermatology, University Hospital of Zurich, Switzerland.

Minor nail trauma may cause bluish discoloration of the nail, while tangential
skin trauma on the heel can result in a so-called black heel. To rule out
melanoma in such clinical situations, a biopsy is needed to reveal homogeneous
eosinophilic masses deposited under the nail plate or within it (transepidermal
elimination). Most dermatopathologists attempt to demonstrate the presence of
hemoglobin in these eosinophilic masses with Prussian blue stain, which typically
remains negative. In our experience, these traumatically induced blood deposits
are always situated in avascular spaces, devoid of degrading phagocytes.
Consequently, a histochemical stain for these deposits should be directed
specifically toward hemoglobin, not hemosiderin. In the dermatopathologic
literature, the various techniques to detect hemoglobin deposits in tissue
sections are not well-known. We would like to emphasize benzidine stain, a highly
selective and efficient method to demonstrate the presence of hemoglobin deposits
in histologic sections. To date, benzidine stain has not been utilized to
characterize splinter hemorrhage (subungual hematoma). Of concern, the use of
benzidine in histopathology laboratories is restricted because this agent is a
known carcinogen, while the non-mutagenic derivative,
3,3',5,5'-tetramethylbenzidine, does not stain histologic sections. Patent blue
V, a completely different and less specific agent, stains hemoglobin an intense
blue-green.





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