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Updated: May 27, 2025
Model of Ischemic Heart Disease and Video-Based Comparison of Cardiomyocyte Contraction Using hiPSC-Derived Cardiomyocytes
Published on: May 5, 2020
Fluorescence in situ hybridization protocol for cardiomyocytes
Zehao Yao1, Lina Bai2, Yu Nie3
1State Key Laboratory of Cardiovascular Disease, Fuwai Hospital, National Center for Cardiovascular Disease, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100037, China; 4+4 Medical Doctor Program, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
We developed cardioFISH, a novel method for measuring cardiomyocyte nuclear ploidy. This technique overcomes cellular complexities, enabling accurate ploidy assessment for cardiac research.
Area of Science:
- Cardiovascular Biology
- Cell Biology
- Genetics
Background:
- Accurate measurement of cardiomyocyte nuclear ploidy is essential for understanding cardiac development, function, and disease.
- Current methods like fluorescence in situ hybridization (FISH) face challenges in cardiomyocytes due to cellular complexity.
Purpose of the Study:
- To present a detailed, cardiomyocyte-specific FISH (cardioFISH) protocol.
- To enable robust and accurate nuclear ploidy assessment in cardiomyocytes.
Main Methods:
- Developed a tailored enzymatic digestion strategy to improve nuclear accessibility and preserve cellular integrity.
- Minimized sarcomere-derived autofluorescence.
- Introduced a 3D nuclear visualization framework for comprehensive signal analysis in large cardiomyocyte nuclei.
Main Results:
- The cardioFISH protocol is a two-day procedure.
- It is applicable across various cardiomyocyte developmental stages.
- The method enhances nuclear accessibility and minimizes autofluorescence.
Conclusions:
- CardioFISH provides a powerful tool for advancing studies on cardiomyocyte nuclear ploidy.
- This protocol addresses limitations of traditional FISH in cardiomyocytes.
- It facilitates comprehensive analysis of cardiomyocyte ploidy for cardiac research.
