Number one - get rid of B5; use formalin!

Number two - you need to use different antibody concentrations and retrieval methods to see light chain-restricted immunoreactivity in formalin-fixed, paraffin-embedded HemePath specimens.

We have been moderately successful using a 96 degree waterbath for 40 minutes for retrieval; however, with lymphomas with abundant cytoplasmic immunoglobulin this method may be too sensitive and you may have to turn to an enzyme digestion (we use pepsin).  We also have two different dilutions for both kappa and lambda.  If the first attempt shows too much background, we will then repeat the studies using lower concentrations of antibody.  Everyone's tissue is different, but this has worked for us.  See our article, "Immunohistochemical detection of immunoglobulin light chain expression in ........" that appeared in Applied Immunohistochemistry & Molecular Morphology back in 2002 (volume 10/number 3).

Richard  

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

>>> Histology SLU  08/23/05 12:53PM >>>
Hello All:
 
We have, for some time now, had issues with our bone marrow core biopsies and staining with Kappa and Lambda.  Our cores are B5 fixed for 2 hours then decaled in RDO for one hour.  They are processed overnight on the tissue processor and embedded the next morning.  If anyone has any other protocols or more specifically could share your handling schedules and K/L protocols, I would be forever grateful.  I am at a dead end and have a pathologist very upset that I can not get this "fixed".  Thanks always for your help.
 
Susan

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