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Robust 3D DNA FISH Using Directly Labeled Probes
Published on: August 15, 2013
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A high-throughput DNA FISH protocol to visualize genome regions in human cells
Elizabeth H Finn1, Tom Misteli1
1Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MA, 20892, USA.
STAR Protocols
|August 30, 2021
Summary
This study presents a rapid, high-throughput imaging protocol for visualizing genomic loci in cells. The method uses DNA fluorescence in situ hybridization and automated microscopy to quantify gene positioning and cellular heterogeneity.
Area of Science:
- Molecular Biology
- Cell Biology
- Genomics
Background:
- Understanding gene positioning is crucial for cell function.
- Quantifying heterogeneity in gene loci requires efficient methods.
Purpose of the Study:
- To develop and present an end-to-end high-throughput imaging protocol.
- To enable visualization and quantification of genomic loci in cells.
Main Methods:
- DNA fluorescence in situ hybridization (FISH)
- Automated microscopy
- Computational analysis
- High-throughput imaging
Main Results:
- A one-week protocol for visualizing genomic loci.
- Method for quantifying patterns of heterogeneity in gene positioning.
- Applicable to subpopulations of cells.
Conclusions:
- The protocol offers a robust solution for high-throughput genomic locus visualization.
- Ensures data quality and provides practical solutions for common experimental challenges.
