Tuesday, December 27, 2005

SUSPECTED LYMPHOMA- PROCESSING

Lymphoproliferative Lesions:
Lymphoproliferative lesions as a class constitute a significant diagnostic challenge to the hematopathologist, and routine preparations are often difficult to adequately characterize. Accordingly, any lesion suspected on clinical grounds to be a lymphoma, "pseudolymphoma", or lymphoproliferative lesion must be processed under supervision of the eye pathology attending. In general, the specimens will be sent to the hematopathology service but check.
Prioritize tissue collection for lymphomas as follows:
1. In the OR, wrap the tissue in gauze soaked in phosphate buffered saline (PBS) over wet ice. Do not immerse in buffer!
2. Tissue for routine paraffin embedment - In the lab, fix a thin slice in neutral buffered formalin, B-5 or paraformaldahyde.
3. Touch preps - Prepare 2 air-dried, unstained glass slides for later fixation and staining with May/Grunwald/Geimsa (MGG) by gently touching the freshly cut surface of the tumor on 3-4 spots on the slide. 2 more slides, prepared the same way, should be rapidly fixed in 95% ethanol for staining with H & E or PAP.
4. Immunohistochemistry / Gene rearrangement studies - Select a large piece of fresh tissue. Partially fill the well of a plastic blocking mold with O.C.T. compound (third drawer under the autopsy eyes). Place the specimen in the well and then fill the well with O.C.T. compound. Immediately place it in 95% ethanol (previously cooled to near freezing by dissolving dry ice in it) and place over a block of dry ice. Fill out the immunohistochemistry request form/These are then sent to the Immunochemistry lab in main pathology, CHS 13-240, x50643. Gene rearrangement studies are sent to the molecular biology lab 332B CHS, x55648 or x65294.
5. Electron microscopy --- Dice a small bit of tissue and place it in Formaldehyde-Glutaraldehyde fixative (2% each). Hold tissue in fixative until H & E sections from #1 above are done.
6. Flow Cytometry --- Fresh tissue is submitted for flow cytometry in a special culture media prepared by Flow Cytometry in Room AL 154, x55806 (x56877). Call them to obtain appropriate media and requistion forms. The specimen must be delivered promptly.
Orders for Immunohistochemistry: Our routine for MALT and orbital lymphoma includes antibodies to: CD3,CD5,CD10,CD20, CD43, BCl2, BCL1, kappa, lambda, BCL6, and IgM. Additional orders per attending.


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