A method of quantifying centrosomes at the single-cell level in human normal and cancer tissue

Mengdie Wang, Beatrice S. Knudsen, Raymond B. Nagle, Gregory C. Rogers, Anne E. Cress

Research output: Contribution to journalArticle

3 Scopus citations

Abstract

Centrosome abnormalities are emerging hallmarks of cancer. The overproduction of centrosomes (known as centrosome amplification) has been reported in a variety of cancers and is currently being explored as a promising target for therapy. However, to understand different types of centrosome abnormalities and their impact on centrosome function during tumor progression, as well as to identify tumor subtypes that would respond to the targeting of a centrosome abnormality, a reliable method for accurately quantifying centrosomes in human tissue samples is needed. Here, we established a method of quantifying centrosomes at a single-cell level in different types of human tissue samples. We tested multiple anti-centriole and pericentriolar-material antibodies to identify bona fide centrosomes and multiplexed these with cell border markers to identify individual cells within the tissue. High-resolution microscopy was used to generate multiple Z-section images, allowing us to acquire whole cell volumes in which to scan for centrosomes. The normal cells within the tissue serve as internal positive controls. Our method provides a simple, accurate way to distinguish alterations in centrosome numbers at the level of single cells.

Original languageEnglish (US)
Pages (from-to)811-819
Number of pages9
JournalMolecular biology of the cell
Volume30
Issue number7
DOIs
StatePublished - Jan 1 2019

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ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

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