[Histonet] Gram Staining

[Tony Henwood (SCHN)]

Check the age of the Iodine.

Iodine solutions lose their activity over time (?notice the brown discolouration on the parafilm sealing a coplin jar) even though the solution still looks as brown as when it was made.

Magee, C. M., Rodeheaver, G., Edgerton, M. T., & Edlich, R. F. (1975). A more reliable gram staining technic for diagnosis of surgical infections. The American Journal of Surgery, 130(3), 341-346.

"Opening the bottle in a warm room potentiated the loss of iodine so rapidly that no iodine could be detected in the solutions after fifteen days. The potentiation of loss of iodine by temperature elevation becomes a severe problem in the warm seasons of the year, particularly in tropical areas." And "When iodine concentration of the solution was less than 20 per cent of the original value, some of the stained slides revealed only gram-negative organisms."


Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Principal Scientist, the Children's Hospital at Westmead
Adjunct Fellow, School of Medicine, University of Western Sydney 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
Pathology Department
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 



-----Original Message-----
From: Ken M via Histonet [mailto:histonet at lists.utsouthwestern.edu] 
Sent: Wednesday, 1 July 2020 7:21 AM
To: histonet at lists.utsouthwestern.edu
Subject: [Histonet] Gram Staining

Hi.

We are trying to stain a supposed Gram Positive paraffin Tonsil tissue block using Gentian Violet, Iodine (Lugol), Gram's decolorizer, Carbol Fuchsin Counter and Tartrazine and are having trouble distinguishing the positive from the negative. Anyone have any suggestions?


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