Immunofluorescence-based Determination of Centrosome Number in Tissue Samples

Area of Science:

• Cell Biology
• Cancer Research
• Microscopy

Background:

• Centrosome numerical abnormalities are common in various tumors, exhibiting significant heterogeneity.
• Centrosome amplification, the overproduction of centrosomes, is a unique characteristic of cancer cells and a potential therapeutic target.

Purpose of the Study:

• To develop and describe a reliable protocol for quantifying centrosome numbers at the single-cell level.
• To enable accurate assessment of centrosome abnormalities in formalin-fixed paraffin-embedded (FFPE) tissue samples.

Main Methods:

• Utilizing multiplexing antibodies to identify bona fide centrosomes and cell borders.
• Employing high-resolution immunofluorescent microscopy with Z-sectioning for single-cell analysis.
• Applying the protocol to diverse human tissue samples.

Main Results:

• Successfully quantified centrosome numbers on a single-cell level in FFPE tissue samples.
• Demonstrated the protocol's ease of use and applicability across various human tissues.
• Provided a method to address the need for single-cell centrosome quantification in cancer research.

Conclusions:

• The described protocol offers a straightforward and effective method for quantifying centrosome numbers in FFPE tissues.
• This technique facilitates the study of centrosome amplification and numerical abnormalities in cancer.
• The protocol supports the advancement of centrosome-targeting cancer therapies.