[Histonet] Podoplanin D2-40

[Richard Cartun]

Are your fluids initially fixed in alcohol before the cell block is prepared?  If so, that can reduce immunoreactivity for some proteins.  If you can, I would recommend trying to decrease the sensitivity of your antigen retrieval.  I am a strong proponent of "Personalized" antigen retrieval (adjusting the retrieval based on the type of specimen, fixation, and the length of fixation).  I do not have any experience with Ventana's instruments, but on our Bond Max and Bond III instruments, we can adjust the antigen retrieval very easily (low vs. high pH and the time).

Richard

Richard W. Cartun, Ph.D.
Director, Histology & Immunopathology
Director, Biospecimen Collection Programs
Assistant Director, Anatomic Pathology
Hartford Hospital
80 Seymour Street
Hartford, CT  06102
(860) 545-1596
(860) 545-0174 Fax

>>> Carol Bryant <cbrya <@t> lexclin.com> 1/11/2010 2:10 PM >>>
I am using Podoplanin, D2-40 on cytology cases.  We use a mesothelioma tissue for the control and the control performs appropriately, but our cell blocks are staining very faintly.  We stain on the Ventana Benchmark using the following protocol:  Depar, standard CC1, antibody for 32 min, amplify selected, counterstain, and post counterstain.   I am thinking the standard CC1 may be too harsh for the cell blocks and maybe cutting it back to mild treatment may help. Any other suggestions to boost the staining intensity of the podoplanin on the cell blocks?
Thank you in advance for your thoughts and input.

Carol

Carol Bryant, CT (ASCP)
Cytology/Histology Manager
Pathology Services
Lexington Clinic
Phone (859) 258-4082
Fax (859) 258-4081
cbrya <@t> lexclin.com 



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