Immunoglobulin light chain staining in paraffin-embedded tissue using a heat mediated epitope retrieval method.
We have compared light chain immunohistochemistry in reactive lymphoid tissue and a series of paraffin-embedded B-cell lymphomas using standard trypsin digestion with a heat mediated epitope retrieval method. Fifty-seven B-cell lymphomas (18 high grade, 29 low grade and 10 cases of nodular lymphocyte predominance Hodgkin's disease), two reactive lymph nodes and eight tonsils fixed for known times between 12 h and 2 years were studied. Paraffin-embedded tissue was stained with polyclonal anti-kappa and anti-lambda antibodies. For each antibody staining was performed on two sections, one treated with trypsin digestion and one with microwave heating. Sections were scored from 0 to + + + with 0 representing poor staining and + + + excellent staining. A score of ++ was considered satisfactory. Light chain restriction was recorded if present. Satisfactory staining was obtained in 34/59 cases using trypsin digestion and 56/59 cases using heat retrieval. Light chain restriction was demonstrated in 32/57 (56%) B-cell lymphomas using trypsin digestion and 52/57 (91%) using heat retrieval. Satisfactory staining was obtained in tonsils fixed for up to 48 h using trypsin digestion and up to 2 years using heat retrieval. We have shown that for light chain immunostaining a heat mediated epitope retrieval method produces more consistent and satisfactory results and is effective over a greater range of fixation times than traditional trypsin digestion.
Ashton-Key
M
M
Department of Histopathology, University College London Medical School, UK.
Jessup
E
E
Isaacson
P G
PG
D000939:Epitopes; D006358:Hot Temperature; D006801:Humans; D007147:Immunoglobulin Light Chains / Q000032:analysis*; D007150:Immunohistochemistry / Q000379:methods*; D008221:Lymphoid Tissue / Q000737:chemistry* / Q000473:pathology; D016393:Lymphoma, B-Cell / Q000737:chemistry* / Q000473:pathology; D008872:Microwaves; D016612:Paraffin Embedding; D013194:Staining and Labeling; D016707:Tissue Fixation
D000939:Epitopes; D007147:Immunoglobulin Light Chains