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A quantitative method for measurement of HL-60 cell apoptosis based on diffraction imaging flow cytometry technique
Xu Yang1, Yuanming Feng2, Yahui Liu1
1Department of Biomedical Engineering, Tianjin University, Tianjin 300072, China.
Biomedical Optics Express
|July 30, 2014
Summary
This study introduces a novel, staining-free method using polarization diffraction imaging flow cytometry to quantify apoptosis in HL-60 cells. Four gray level co-occurrence matrix parameters accurately measure apoptotic rates and nuclear fragmentation.
Area of Science:
- Cell biology
- Biophysics
- Quantitative imaging
Background:
- Apoptosis quantification is crucial for understanding cell death.
- Existing methods often require cell staining, which can be time-consuming and introduce artifacts.
- Developing rapid, accurate, and staining-free methods for apoptosis measurement is highly desirable.
Purpose of the Study:
- To present a quantitative method for measuring apoptosis in HL-60 cells.
- To utilize polarization diffraction imaging flow cytometry combined with gray level co-occurrence matrix (GLCM) analysis.
- To assess the sensitivity and correlation of GLCM parameters with apoptotic rates and nuclear morphology.
Main Methods:
- Employed polarization diffraction imaging flow cytometry for cellular analysis.
- Applied gray level co-occurrence matrix (GLCM) algorithm to analyze diffraction images.
- Investigated four GLCM parameters: contrast (CON), cluster shade (CLS), correlation (COR), and dissimilarity (DIS).
- Compared the new method with existing apoptosis measurement techniques.
Main Results:
- Four GLCM parameters (CON, CLS, COR, DIS) demonstrated high sensitivity in detecting apoptotic rates.
- The CLS parameter showed a significant correlation (R(2) = 0.899) with nuclear fragmentation.
- The other three parameters exhibited very good correlations (R(2) ranging from 0.69 to 0.90) with apoptotic indicators.
- The method successfully quantified cellular apoptosis without requiring cell staining.
Conclusions:
- The developed polarization diffraction imaging flow cytometry method offers a rapid and accurate approach for apoptosis quantification.
- GLCM analysis of diffraction images provides sensitive morphological features for assessing apoptosis.
- This staining-free technique eliminates the need for cell labeling, simplifying the apoptosis measurement process.
