Double labeling and simultaneous detection of B- and T cells using fluorescent nano-crystal (q-dots) in paraffin-embedded tissues.
A double immunohistochemical technique for the simultaneous detection of T- and B cells in paraffin-embedded mice tissues have been developed. This procedure is based on using fluorescent nano-crystals (q-dots). The benefit of using q-dots evolves from their unique fluorescence characteristics advantages: such as broad excitation spectrum, narrow emission band and high photo-bleaching threshold compare to organic fluorophores. T cells antigens (CD3) were stained using antibody-coated q-dots with max emission at 655 nm (GalphaRb-QD655). B cells antigens (CD45R/B220) were stained using streptavidin-coated q-dots with max emission at 585 nm (SA-QD585). The simultaneous detection of T- and B cells was demonstrated in paraffin-embedded lymph node using standard fluorescence microscope.
Zahavy
Eran
E
Department of Infectious Diseases, Israel Institute for Biological Research, Ness-Ziona, 74100, Israel.
Freeman
Esther
E
Lustig
Shlomo
S
Keysary
Avi
A
Yitzhaki
Shmuel
S
D000818:Animals; D000906:Antibodies / Q000737:chemistry; D001402:B-Lymphocytes / Q000276:immunology*; D017252:CD3 Complex / Q000032:analysis* / Q000737:chemistry; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes / Q000737:chemistry*; D017493:Leukocyte Common Antigens / Q000032:analysis* / Q000737:chemistry; D051379:Mice; D008856:Microscopy, Fluorescence; D049329:Nanostructures / Q000737:chemistry; D016612:Paraffin Embedding; D010777:Photochemistry; D045663:Quantum Dots; D013050:Spectrometry, Fluorescence; D013194:Staining and Labeling; D019809:Streptavidin / Q000737:chemistry; D013601:T-Lymphocytes / Q000276:immunology*; D016610:Tissue Embedding
D000906:Antibodies; D017252:CD3 Complex; D005456:Fluorescent Dyes; D019809:Streptavidin; D017493:Leukocyte Common Antigens