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Clonality analysis of defined B‐cell populations in archival tissue sections using microdissection and the polymerase chain reaction
Author(s) -
PAN L.X.,
DISS T.C.,
PENG H.Z.,
ISAACSON P.G.
Publication year - 1994
Publication title -
histopathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.626
H-Index - 124
eISSN - 1365-2559
pISSN - 0309-0167
DOI - 10.1111/j.1365-2559.1994.tb00532.x
Subject(s) - microdissection , laser capture microdissection , polymerase chain reaction , polyclonal antibodies , microbiology and biotechnology , biology , pathology , lymphoma , b cell , antibody , gene , immunology , medicine , genetics , gene expression
A simple microdissection technique involving the use of a drawn‐out glass pipette was developed for isolation of defined cell subsets from tissue sections. Using this technique and the polymerase chain reaction (PCR), clonally rearranged immunoglobulin (Ig) heavy chain genes were reliably amplified in single neoplastic follicles or few hundreds of tumour cells isolated from archival haematoxylin and eosin or immunostained sections of B‐cell lymphomas. A polyclonal nature was consistently demonstrated in reactive lymphoid follicles or interfollicular reactive B‐cells within the same lymphoma sections. Microdissection of lymphoma cells from within foci of chronic inflammation improved the resolution of tumour‐specific PCR products by reducing amplification of background polyclonal B‐cell sequences. The combination of microdissection and PCR techniques, therefore, provides an important tool for the investigation of B‐cell lymphomas and also allows simple and specific access for other molecular genetic analyses of different cell subsets on tissue sections.