Identification of leucocyte surface antigens in paraffin-embedded bovine tissues using a modified formalin dichromate fixation.
A modified fixative of formalin dichromate was combined with a cold embedding procedure for the preservation of bovine leucocyte surface antigens. Fourteen monoclonal antibodies recognizing seven bovine leucocyte surface antigens (BoCD1w2, BoCD4, BoCD8, BoWC1, BoWC3, BoWC4 and BoIgM) were applied as primary antisera in a sensitive avidin-biotin-peroxidase complex detection method. The staining results were compared with those obtained in cryostat and routinely formalin-fixed sections of corresponding tissue samples. Using the modified formalin dichromate fixative and the cold embedding procedure, all the leucocyte surface antigens tested were detectable immunohistologically in paraffin sections with a generally more distinct staining than in traditionally processed tissues. Morphological structures were better preserved than in cryostat sections but, to some extent, were poorer when compared with routinely formalinfixed tissues. However, this method suggests that there are only mild masking effects and provides an alternative to the use of unfixed material, particularly for morphological-immunohistochemical investigations.
Rathkolb
B
B
Institut für Pathologie, Tierärztliche Hochschule Hannover, Germany.
Pohlenz
J F
JF
Wohlsein
P
P
D000818:Animals; D018949:Antigens, CD1 / Q000032:analysis; D000954:Antigens, Surface / Q000032:analysis*; D015704:CD4 Antigens / Q000032:analysis; D016827:CD8 Antigens / Q000032:analysis; D002417:Cattle; D002840:Chromates / Q000737:chemistry*; D005557:Formaldehyde / Q000737:chemistry*; D007075:Immunoglobulin M / Q000032:analysis; D007150:Immunohistochemistry; D007962:Leukocytes / Q000737:chemistry; D008198:Lymph Nodes / Q000737:chemistry; D008221:Lymphoid Tissue / Q000737:chemistry*; D008562:Membrane Glycoproteins / Q000032:analysis; D014066:Palatine Tonsil / Q000737:chemistry; D016612:Paraffin Embedding / Q000379:methods; D010581:Peyer's Patches / Q000737:chemistry; D013154:Spleen / Q000737:chemistry; D016707:Tissue Fixation / Q000379:methods*
D018949:Antigens, CD1; D000954:Antigens, Surface; D015704:CD4 Antigens; D016827:CD8 Antigens; D002840:Chromates; C086500:DMBT1 protein, Bos taurus; D007075:Immunoglobulin M; D008562:Membrane Glycoproteins; D005557:Formaldehyde; C034944:chromic acid