Comparison of labeling methods and embedding media for localization of S-phase osteogenic cells

Caroline Jennermann, Rosario H. Potter, Lawrence P. Garetto

Research output: Contribution to journalArticle

Abstract

Non-radioactive 5-bromo-2'-deoxyuridine (BrdU) immunohistochemistry has been established as a substitute for 3H-thymidine autoradiography in localizing cells undergoing DNA synthesis (S-phase). This experiment compared the 2 methods using an in vivo model for osteoblast histogenesis that separates cells into 2 proliferating precursor compartments. Twenty Sprague Dawley rats were equally divided with half given 3H-thymidine (1 μCi/gm) and half given BrdU (50 μgm/gm). Mandibular condyles were embedded (one side in paraffin, the other in methyl methacrylate) and S-phase cells identified using either thymidine autoradiography or BrdU immunohistochemistry. The fractional distribution of osteoblast precursor cell types for each specimen and the percent of labeled cells (within each cell type and label index of randomly selected cells) were calculated. Chi-square analysis showed no difference between embedding or labeling methods in the overall cell distribution. However, when cells were separated into kinetic cell types, significant differences in percent labeling were shown by analysis of variance. These results suggest that the 2 labeling methods do not yield the same results in osteogenic tissue when cells are divided into kinetic subpopulations. Furthermore, embedding media is a critical factor in the effectiveness of the BrdU labeling method.

Original languageEnglish (US)
Pages (from-to)39-44
Number of pages6
JournalJournal of Histotechnology
Volume20
Issue number1
StatePublished - 1997
Externally publishedYes

Fingerprint

S Phase
Bromodeoxyuridine
Thymidine
Autoradiography
Osteoblasts
Immunohistochemistry
Mandibular Condyle
Methacrylates
Paraffin
Sprague Dawley Rats
Analysis of Variance
DNA

Keywords

  • 5-bromo-2'-deoxyuridine
  • H- thymidine
  • autoradiography
  • embedding media
  • immunohistochemistry
  • osteogenesis

ASJC Scopus subject areas

  • Anatomy
  • Cell Biology

Cite this

Comparison of labeling methods and embedding media for localization of S-phase osteogenic cells. / Jennermann, Caroline; Potter, Rosario H.; Garetto, Lawrence P.

In: Journal of Histotechnology, Vol. 20, No. 1, 1997, p. 39-44.

Research output: Contribution to journalArticle

Jennermann, Caroline ; Potter, Rosario H. ; Garetto, Lawrence P. / Comparison of labeling methods and embedding media for localization of S-phase osteogenic cells. In: Journal of Histotechnology. 1997 ; Vol. 20, No. 1. pp. 39-44.
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