A method to quantify centrosomes at single-cell level in human normal and cancer tissue.

Centrosome abnormalities are emerging hallmarks of cancer. The over-production 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 identify tumor subtypes that would respond to targeting a centrosome abnormality, a reliable method to accurately quantify centrosomes in human tissue samples is needed. Here, we established a method to quantify 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 with 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.