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

1 Citation (Scopus)

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 - Mar 21 2019

Fingerprint

Centrosome
Neoplasms
Centrioles
Cell Size
Microscopy

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

A method of quantifying centrosomes at the single-cell level in human normal and cancer tissue. / Wang, Mengdie; Knudsen, Beatrice S.; Nagle, Raymond B; Rogers, Gregory C.; Cress, Anne E.

In: Molecular biology of the cell, Vol. 30, No. 7, 21.03.2019, p. 811-819.

Research output: Contribution to journalArticle

@article{8ee8b6d2a4744da1b0885ebd6156e2f4,
title = "A method of quantifying centrosomes at the single-cell level in human normal and cancer tissue",
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.",
author = "Mengdie Wang and Knudsen, {Beatrice S.} and Nagle, {Raymond B} and Rogers, {Gregory C.} and Cress, {Anne E}",
year = "2019",
month = "3",
day = "21",
doi = "10.1091/mbc.E18-10-0651",
language = "English (US)",
volume = "30",
pages = "811--819",
journal = "Molecular Biology of the Cell",
issn = "1059-1524",
publisher = "American Society for Cell Biology",
number = "7",

}

TY - JOUR

T1 - A method of quantifying centrosomes at the single-cell level in human normal and cancer tissue

AU - Wang, Mengdie

AU - Knudsen, Beatrice S.

AU - Nagle, Raymond B

AU - Rogers, Gregory C.

AU - Cress, Anne E

PY - 2019/3/21

Y1 - 2019/3/21

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85063606762&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85063606762&partnerID=8YFLogxK

U2 - 10.1091/mbc.E18-10-0651

DO - 10.1091/mbc.E18-10-0651

M3 - Article

C2 - 30699045

AN - SCOPUS:85063606762

VL - 30

SP - 811

EP - 819

JO - Molecular Biology of the Cell

JF - Molecular Biology of the Cell

SN - 1059-1524

IS - 7

ER -