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[Continual and quantitative analysis of cellular apoptosis by electron and fluorescence microscope]
Shenglan Wang1, Yanfeng Zhong, Zengwei Guan
1Department of Pathology, Peking University School of Basic Medical Sciences, Beijing 100083, China.
Summary
Fluorescent microscopy effectively analyzes cellular apoptosis and necrosis by distinguishing morphological changes. This technique provides a quantitative method for observing programmed cell death progression over time.
Area of Science:
- Cell Biology
- Microscopy Techniques
Context:
- Cellular apoptosis and necrosis are critical biological processes.
- Distinguishing between apoptosis and necrosis is crucial for understanding cell fate.
Purpose:
- To evaluate the utility of fluorescent microscopy for analyzing cellular apoptosis.
- To compare fluorescent microscopy with transmission electron microscopy in apoptosis studies.
Summary:
- VP16 and Sodium Azide (NaN3) were used to induce apoptosis and necrosis in HL60 cells, respectively.
- Fluorescent microscopy, using Hoechst 33258 staining, revealed distinct morphological features: pyknosis, chromatin aggregation, and karyorrhexis in apoptosis, versus karyolysis and chromatin diffusion in necrosis.
- Time-course analysis demonstrated that pyknosis precedes karyorrhexis during apoptosis, with pyknosis observed at 4 hours and peaking at 8 hours, and karyorrhexis reaching 80% by 24 hours.
Impact:
- Fluorescent microscopy offers a continual and quantitative method for apoptosis analysis.
- This technique can reliably differentiate between cellular apoptosis and necrosis.
- The findings support the use of fluorescent microscopy as a valuable tool in cell death research.