Detection of T‐cell receptor β chain mRNA in frozen and paraffin‐embedded biopsy tissue using digoxigenin‐labelled oligonucleotide probes in situ

Abstract

In situ hybridization techniques using a cocktail of digoxigenin-labelled T-cell receptor (TcR) constant (C) region beta oligonucleotide probes were used to detect TcR beta mRNA in frozen and paraffin-embedded tissue sections. The specificity of the C beta cocktail was confirmed by Northern blot analysis. The TcR C beta cocktail successfully hybridized to T cells in frozen and paraffin-embedded tissue obtained from patients with inflammatory arthropathies, B- and T-cell non-Hodgkin's lymphoma (NHL), and reactive tonsillitis, and showed staining patterns comparable to those obtained by conventional immunohistological detection of T cells. This is the first report of in situ studies using labelled TcR C beta oligonucleotide probes and may indicate the feasibility of investigating clonal T-cell populations using digoxigenin-labelled clonospecific probes in clinical samples in situ.

Isolating RNA from clinical samples with catrimox-14 and lithium chloride

“…nohistochemical staining. We found reports of in situ identification of mRNA in frozen sections (18,19), but we have not found reports of their use as a source of clinical material for RNA isolation. We found that RNA can be isolated from frozen sections with Catrimox or the GPC reagent.…”