[Histonet] Inconsistent immuno staining

[Tony Henwood]

Ann,

I assume you are demonstrating mast cells. Mast cells are very small cells and on sectioning it is easy to only have less than half a mast cell in your sections, where as you will find whole mast cells close by.
This difference will mirror your staining: strong staining in the whole cells, weaker staining in the sectioned cells. You will see the same occurring with the mast cell metachromatic stains (toluidine blue or Giemsa), strong and weaker staining depending on whether you have whole or sectioned cells.

Does this help?

Regards 
Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC), FFSc(RCPA) 
Laboratory Manager & Senior Scientist 
Tel: 612 9845 3306 
Fax: 612 9845 3318 
the children's hospital at westmead
Cnr Hawkesbury Road and Hainsworth Street, Westmead
Locked Bag 4001, Westmead NSW 2145, AUSTRALIA 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Ann Bennett
Sent: Thursday, 13 January 2011 10:37 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Inconsistent immuno staining

Hello Histonetters,
 
While trying to eliminate any variables - processing times, embedding paraffin, waterbath temperature, time and temp in oven, deparaffinization, antigen retrieval, antibody manufacturer and lot # - I still seem to have a slight inconsistency with 1 of my immuno stains.
 
Mast Cell Tryptase staining for colon biopsies.
 
Can anyone offer any suggestions or ideas as to why this might be occuring?
 
Even within my current control slides I see a nice distinct brown or a slightly more pale stain.
 
I would love to hear your ideas.
 
Thank you!
 
Ann.BennettAnn


      
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