[Histonet] JB Fixative

[Gayle Callis]

Is there someone selling this fixative under the name, JB fixative?  After 
Jay Beckstead, this fixative was later used by

Nitta et al, Cell Vision, Improved in situ immunodetection of leukocytes on 
paraffin embedded mouse spleen 4(1):73-80, 1997
  (a defunct journal) to avoid any kind of formalin fixation for murine CD 
markers.

Nitta never called it JB, but I really like this acronym/name!!!   This is 
an excellent publication to read as it also refers to antibodies they use, 
vendor, along with tissue processing and IHC method used.  We repeated IHC 
results nicely using Alk Phos IHC method.

I have not seen this fixative referred to as JB fixative.  BD Pharmingen 
refers to and sells it under the name Zinc Fixative but do us (or have done 
in the past) no favors with their name for this "JB"  fixative.  People 
tend to immediately confuse it with Zinc formalin - a case of similar names 
but very dissimilar fixatives, particularly when formalin is NOT wanted for 
these murine CD markers.   Maybe BD Pharmingen is looking in, and will do 
us a favor with a distinct renaming - JB fixative!!   It is still called 
Zinc fixative in their online catalog although they did add "formalin 
free".  Ho Hum!

Our lab always called it ZincTRIS fixative (ZnTRIS for short) to 
(hopefully) not confuse it with zinc formalin, and I plan to change it to 
JB - so tidy!

***Nitta indicated fixation of murine tissues could be as long as 72 hours 
- rather than just 48 hours.
Although JB/PE tissues are touted to give tissue morphology comparable to 
FFPE tissues the fixative can present some crunchy tissue blocks, rather 
dry and friable noticed at sectioning. We had to do long ice water soaking 
before sectioning.  It gives adequate morphology but not comparable per 
previous comment from our experience.  Also, be careful about placing too 
large a tissue in this fixative.  If the tissue is NOT totally fixed by JB, 
then 70% alcohol and processing alcohols will finish the fixation for you, 
not ideal for IHC purposes - this happened to a colleague of mine when she 
tried JB fixative.

Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610






  At 07:37 AM 3/24/2005, you wrote:
>Megan,
>Here is the "recipe" for the JB fixative which we have used for fixation
>of mouse tissues in which we wanted to detect CD3, CD4 and CD8.
>
>JB Fixative
>
>0.1 M  Tris Buffer, pH 7.4              1000ml
>Calcium acetate                       0.5 g
>Zinc Acetate                            5.0 g
>Zinc Chloride                           5.0 g
>
>Mix to dissolve.  The final pH will be approximately 6.8 (6.5-7.0).  Do
>not readjust the pH, as this will cause the zinc to come out of
>solution.  Small amounts of insoluable zinc oxychloride often
>contaminates zinc chloride.  This may result in a very small amount of
>white precipitate.  This will not cause any problems but can be removed
>by filtering the solution.
>
>Fix tissue for 24-48 hours, then transfer tissues to 70% ethanol.
>
>Reference:
>Beckstead JH. A Simple Technique for Preservation of Fixative-sensitive
>Antigens in Paraffin-embedded Tissues: Addendum (Letter to the Editor).
>J Histochem Cytochem 1994; 43:345
>
>
>Pat Greer
>Centers for Disease Control and Prevention
>Infectious Disease Pathology Activity
>1600 Clifton Road, MS G-32
>Atlanta, GA 30333
>
>
>I require some information regarding these fixatives. If anyone out in
>Histonetland is is using any of these fixatives or has a reference for
>me , I would be very happy and thankful.
>
>CBA formalin
>JB fixative
>Excell
>
>Thank you for your help in advance with my query.
>
>HAPPY EASTER and HAPPY HOLIDAYS
>
>Zenobia Haffajee
>Hunter Area New England Health
>IHC dept
>Newcastle
>Australia
>
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Gayle Callis
MT,HT,HTL(ASCP)
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367 (lab with voice mail)
406 994-4303 (FAX)