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- Title
Rapid immunofluorescent determination of cells in the S phase in pea root meristems: An alternative to autoradiography.
- Authors
Levi, Marisa; Sparvoli, Elio; Sgorbati, Sergio; Chiatante, Donato
- Abstract
Root meristems of Pisum sativum L. cv. Lincoln were pulsed with bromodeoxyuridine, and after partial DNA denaturation the incorporated precursor was detected with an indirect immunofluorescent method by means of commercial anti-bromodeoxyuridine monoclonal antibody and fluorescein isothiocyanate-labeled secondary antibody. The nuclei were counterstained with the DNA-specific fluorochrome 4',6-diamidino-2-phenylindole (DAN). The labeling indices (percentage of labeled cells) determined on slides prepared with the same nuclear population were very similar and-in strict accordance with autoradiographic data. When DAPI fluorescence of the nuclei was measured, the unlabeled nuclei coincided with the (ii and part of the G2 regions, whereas all the labeled nuclei were in the S or G2 region. The method is reliable and, in contrast to autoradiography, data can be collected rapidly and almost im - mediately after the pulse. Furthermore, labeling index and the distribution of DNA contents can be obtained from the same slide.
- Subjects
PEAS; BROMODEOXYURIDINE; DNA; IMMUNOFLUORESCENCE; MONOCLONAL antibodies; AUTORADIOGRAPHY
- Publication
Physiologia Plantarum, 1987, Vol 71, Issue 1, p68
- ISSN
0031-9317
- Publication type
Article
- DOI
10.1111/j.1399-3054.1987.tb04618.x