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Updated: May 9, 2026
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Quantitative Immunofluorescence Assay to Measure the Variation in Protein Levels at Centrosomes
Published on: December 20, 2014
Quantitative analysis of centromeric FISH spots during the cell cycle by image cytometry
Genta Amakawa1, Kenzo Ikemoto, Hideaki Ito
1Department of Pathology, Yamaguchi University Graduate School of Medicine, Ube, Japan (GA, KI, HI, TF, KS).
Summary
This study shows that the fluorescence intensity of DNA probes in breast cancer cells increases during the late S and G2 phases of the cell cycle. This finding reveals new insights into cell cycle progression using advanced imaging techniques.
Area of Science:
- Cell Biology
- Genetics
- Cancer Research
Background:
- Fluorescence in situ hybridization (FISH) is a molecular cytogenetic technique.
- Understanding cell cycle progression is crucial for cancer research.
Purpose of the Study:
- To investigate the association between FISH signal intensity and cell cycle progression in breast cancer cells.
- To determine if quantitative analysis of FISH spot intensity can reveal cell cycle dynamics.
Main Methods:
- Two-color FISH using chromosome-specific DNA probes (chromosomes 7, 11, 17, 18) on CAL-51 breast cancer cells.
- Quantitative analysis of FISH spot fluorescence intensity.
- Cell cycle staging using image cytometry at the single-cell level.
Main Results:
- FISH spot intensity was significantly higher in G2 phase cells compared to G0/1 phase cells.
- Increased spot intensity was observed in late S phase, reaching levels similar to G2 phase.
- This indicates alpha satellite DNA replication in the centromeric region during late S phase.
Conclusions:
- Image cytometry can accurately detect subtle differences in centromeric FISH spot intensity.
- Combining FISH analysis with cell image cytometry provides novel insights into cell cycle mechanisms.
- This study is the first to demonstrate the use of image cytometry for analyzing FISH signal intensity throughout the cell cycle.
