BrdUrd labeling of S-phase cells in testes and small intestine of mice, using microwave irradiation for immunogold-silver staining: an immunocytochemical study.
In this study, BrdUrd labeling of S-phase cells in the small intestine and testes was accomplished using microwave irradiation. In this way crypt cells, spermatogonia, and Leydig cells could be labeled using removable plastic-embedded sections and immunogold-silver staining (IGSS). By using short periods of microwave irradiation for incubation of the monoclonal antibodies and the protein A-colloidal gold solution, the detection of BrdUrd-labeled cells could be remarkably enhanced. A comparative study of BrdUrd labeled spermatogonia in the testis of a Cpb-N mouse that received both [3H]-thymidine and BrdUrd proved that 90% of the BrdUrd-labeled cells also showed [3H]-thymidine labeling. The radioactive [3H]-thymidine labeling was a time-consuming method of 4 weeks' duration, whereas the BrdUrd-labeled cells could be labeled, fixed, enhanced, and counterstained in less than 3 hr. This investigation proves that BrdUrd labeling of S-phase cells can be a reliable, reproductive, rapid, and non-radioactive alternative method for [3H]-thymidine labeling of proliferating cells.
Thoolen
B
B
Department of Veterinary Pathology, Veterinary Faculty, State University, Utrecht, The Netherlands.
D000818:Animals; D001973:Bromodeoxyuridine / Q000032:analysis / Q000378:metabolism; D002453:Cell Cycle; D002467:Cell Nucleus / Q000378:metabolism / Q000648:ultrastructure; D007150:Immunohistochemistry; D007421:Intestine, Small / Q000166:cytology* / Q000378:metabolism; D007985:Leydig Cells / Q000166:cytology / Q000378:metabolism; D008297:Male; D051379:Mice; D008859:Microscopy, Polarization; D008872:Microwaves; D012671:Seminiferous Tubules / Q000166:cytology / Q000378:metabolism; D012834:Silver; D013076:Sperm Count; D013094:Spermatozoa / Q000166:cytology / Q000378:metabolism; D013194:Staining and Labeling; D013737:Testis / Q000166:cytology* / Q000378:metabolism; D013936:Thymidine / Q000032:analysis / Q000378:metabolism; D014021:Tissue Preservation
D012834:Silver; D001973:Bromodeoxyuridine; D013936:Thymidine