Dear list,
Those of you who use eosin in your 95% etoh on the processor-what exactly do you use? The ready to stain eosin, or more concentrated? I remember someone writing that they put it in the first 95,correct? We need to stain our tiny gi bxs to better see them when embedding and cutting. Any tips are appreciated.
Jo

>>> "Melanie Black"  10/21/04 02:28AM >>>
>I have also had rod-like crystals in my Alk phos/NBT system. I have 
>shown them to the company rep, who has no idea what they might be. 
>With so much written about it here...I think DAKO should sit up and 
>take a look.

Melanie Black

>
>Thanks all for the cat scratch advice. I am trying it with HRP- AEC. 
>Will report result.
>Jo
>
>
>>>>  "C.M. van der Loos"  10/17/04 02:20PM >>>
>Dear Joanne,
>
>Usually, AP staining doesn't show up in a crystal-like precipitate, 
>but I have seen it a couple of times. Never found out what was the 
>cause of it. Be sure that after your IHC procedure and before the AP 
>visualization, the sections are washed at with a Tris-HCl pH8.5 
>buffer. Phosphate ions from PBS will inhibit your AP reaction. You 
>also may mix the Permanent Red components first and than press 
>immediately through a syringe-filter to remove all bigger particles 
>in the AP solution.
>Have you viewed your slides under a fluorescence microscope? A small 
>amount of Permanent Red reaction product also gives a bright red 
>fluorescence signal when using the rhodamine filter pack (green 
>light).
>Good suggestion by Richard to use a tonsil as a negative control. 
>But, how about running the good old control blanc slide without any 
>primary antibody?
>Hope this helps.
>
>Chris van der Loos, PhD
>Dept. of Pathology
>Academical Medical Center
>Amsterdam - The Netherlands   
>
>----- Original Message -----
>>From  "Joanne Mauger" 
>Date  Fri, 15 Oct 2004 15:45:05 -0400
>To  histonet@lists.utsouthwestern.edu, jkiernan@uwo.ca 
>Subject  Re: [Histonet] IHC withquantum dots
>Hi group,
>Can anyone help me with suggestions-I am working up the monoclonal 
>Ab (H2A10) to cat scratch disease. Ab from Novus Biologicus. I have 
>a purchased control from Newcomer Supply. I am using permanent red 
>chromagen from Dako., with alk phos link also from Dako. I am 
>getting a nice specific staining of what looks like specs vs tiny 
>rods or spirochetes, but the control tissue does , by Warthin 
>Starry, look much more spirochete-ish. Can I trust this result? I 
>have also stained several suspected + cases and see the same dust 
>like + staining,but I am afraid it is teeny bits of precipitate.
>Does this ring a bell?
>Please help.
>Thanks,
>Jo
>
>----- Original Message -----
>>From  "Richard Cartun" 
>Date  Fri, 15 Oct 2004 16:45:57 -0400
>To  ,, 
>
>Subject  Re: [Histonet] IHC withquantum dots
>Have you tried staining a section of tonsil to see if you get the 
>same pattern of staining.  I have found "precipitation" to be a 
>problem with alkaline phosphatase (PA) detection, although I am by 
>no means an expert when it comes to AP IHC.  I use HRP/AEC for most 
>of my infectious disease studies including Bartonella henselae. 
>Don't forget that you may not see the normal morphology of the 
>bacterium with IHC.  You are labeling epitopes on the surface of the 
>organism; you are not staining the entire cell wall.  I have found 
>B. henselae to be one of the most difficult organisms to label (and 
>interpret) with IHC.  Good luck!
>
>Richard
>
>
>Richard W. Cartun, Ph.D.
>Director, Immunopathology & Histology
>Assistant Director, Anatomic Pathology
>Hartford Hospital
>80 Seymour Street
>Hartford, CT  06102
>
>
>
>
>
>
>
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