Re: [Histonet] question

From:"Anthony Reilly"



Kathy
 
You have appeared to try every bluing agent possible, however in my experience the most common cause of pale nuclei in frozen sections is inadequate fixation.  What are you using?  The best I have used is 5% Conc Formalin in 95% Ethanol for at least 30 sec and longer if time permits.
 
All the best.
 
 
 
Tony Reilly

Chief Scientist
Anatomical Pathology
Pathology Queensland
Level 1, Building 15
Princess Alexandra Hospital
Ipswich Rd, 
Woolloongabba Q 4102
Australia
Ph: 07 32402412
Fax:07 32402930
tony_reilly@health.qld.gov.au


>>> "Stephen Peters M.D."  6/06/2008 1:34 am >>>
Kathy,
   
  You may also want to consider the thickness of your sections. A great deal of our information is gatered at scanning magnification of 2x or 4x. If you are cutting at 3 or 
  4 microns or if your cryostat is offering a variety of thick and thin the slides will 
  be pale at these powers compared to 5 or 6 micron sections. Presence of a  lot of 
  nuclear "holes"is a sign it is cut very thin.   Also make sure your hematoxalin is 
  changed regularly and is not growing "rock candy" in the bottom. Check slide after it leaves the bluing and get to know the shade of blue that represents a well stained slide. Remember when checking the slides, the amount of blue will depend on the densityof nuclear material as well as the thickness of the tissue. I find the actual shade 
  of blue tells me that it has been stained in hemotoxalin long enough. 


Stephen Peters M.D. 
Vice Chairman of Pathology
Hackensack University Medical Center 
201 996 4836

Pathology Innovations, LLC 
410 Old Mill Lane, 
Wyckoff, NJ 07481 
201 847 7600 
www.pathologyinnovations.com 




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