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Related Experiment Video

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A High-Throughput In Situ Method for Estimation of Hepatocyte Nuclear Ploidy in Mice

A High-Throughput In Situ Method for Estimation of Hepatocyte Nuclear Ploidy in Mice

Published on: April 19, 2020

High-resolution cell cycle and DNA ploidy analysis in tissue samples.

Christina Heinlein¹, Daniel Speidel^1,2

¹Children's Medical Research Institute, Westmead, New South Wales, Australia.

Current Protocols in Cytometry

|April 2, 2011

Summary

This new method offers a fast, universal way to prepare tissue for cell cycle and DNA ploidy analysis. It avoids harsh chemicals, using a simple mechanical process for high-resolution results.

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Area of Science:

Biotechnology
Cell Biology
Molecular Biology

Background:

Accurate cell cycle and DNA ploidy analysis is crucial for understanding tissue health and disease.
Existing protocols for tissue processing often involve enzymes or detergents, which can be time-consuming and may affect cell integrity.

Purpose of the Study:

To develop an easy, rapid, and universal procedure for processing fresh and frozen tissue specimens.
To enable high-resolution cell cycle and DNA ploidy analysis without harsh chemicals.

Main Methods:

A two-step mechanical tissue dispersion process (approx. 5 minutes).
Single-cell suspension fixation with ethanol and propidium iodide staining.
Flow cytometric DNA content analysis.

Main Results:

The method is applicable to various tissue types (excluding bone) across multiple species.
Achieves highly reproducible cell cycle profiles with excellent resolution.
Reliably detects subtle cell cycle and ploidy alterations, including cell cycle arrest, aneuploidy, and DNA fragmentation.

Conclusions:

This protocol provides a simple, rapid, and versatile approach for tissue processing for cell cycle and DNA ploidy analysis.
It offers a valuable tool for detecting genetic and cellular abnormalities in various biological samples.