The use of FITC-conjugated monoclonal antibodies for determination of S-phase cells with fluorescence microscopy.
A method is modified to determine the DNA synthesizing cells in primary human breast tumors and cells with idiopathic thrombocytopenic purpura (ITP) with FITC-conjugated monoclonal antibody against bromodeoxyuridine (FITC-M-anti-BrdUrd) and fluorescence microscopy. The DNA synthesizing cells were also determined from a portion of the same tissues by classical tritiated thymidine labeling (3HdThd) and autoradiography. The results from bromodeoxyuridine labeling index (BrdUrd-LI) and tritiated thymidine labeling index (3HdThd-LI) obtained from the same tissues were compared. The mean BrdUrd-LI for breast tumor was 5.4 +/- 1.0% and the mean 3HdThd-LI was 5.5 +/- 1.1%. Similarly, the labeling indexes obtained from mononuclear leukocytes of healthy donors had means of 0.5 +/- 0.1% and 0.6 +/- 0.1% for BrdUrd-LI and 3HdThd-LI, respectively. The change in the proliferation rate of mononuclear leukocyte population in the samples obtained from patients with ITP could be observed by both methods. The mean BrdUrd-LI of mononuclear leukocytes for this hematological disorder was 5.4 +/- 0.8%. These results suggest that was 6.1 +/- 0.8%. These results suggest that this relatively simple technique offers an alternative method for determining the DNA synthesizing cells in a given cell population.
D000911:Antibodies, Monoclonal; D001345:Autoradiography; D001943:Breast Neoplasms / Q000473:pathology; D001973:Bromodeoxyuridine / Q000276:immunology; D002453:Cell Cycle; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D005455:Fluorescent Antibody Technique; D006801:Humans; D007399:Interphase; D008214:Lymphocytes / Q000166:cytology; D009000:Monocytes / Q000166:cytology; D011696:Purpura, Thrombocytopenic / Q000473:pathology; D013861:Thiocyanates
D000911:Antibodies, Monoclonal; D005452:Fluoresceins; D013861:Thiocyanates; D001973:Bromodeoxyuridine; D016650:Fluorescein-5-isothiocyanate