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Quantification of apoptotic cells using isolated glomeruli

S Omori1, T Morioka, M Wakasugi

  • 1Department of Cell Biology, Niigata University School of Medicine, Japan.

Nephron
|January 1, 1997
PubMed
Summary
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Detecting kidney cell death is challenging. Combining TUNEL staining with cell markers on isolated glomeruli improves the analysis of cell turnover in kidney disease.

Area of Science:

  • Nephrology
  • Cell Biology
  • Histology

Background:

  • Apoptotic cell detection in kidney disease is crucial for understanding disease progression.
  • The terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end-labeling (TUNEL) method is a standard technique for identifying apoptosis.
  • Current methods using TUNEL on kidney tissue sections yield low detection rates of apoptotic cells per glomerulus.

Purpose of the Study:

  • To develop a more sensitive method for quantifying apoptotic cells within glomeruli.
  • To improve the analysis of cell turnover in experimental nephritis models.

Main Methods:

  • Isolation of glomeruli from kidney tissue.
  • Application of the TUNEL method to whole isolated glomeruli.
  • Immunostaining with cell-specific markers on isolated glomeruli.

Related Experiment Videos

  • Analysis using laser scan microscopy.

    • Main Results:

    • The combined approach of TUNEL and cell-specific marker immunostaining on isolated glomeruli enhances the detection of apoptotic cells.
    • This method allows for more accurate quantification of cell turnover within glomeruli compared to traditional tissue section analysis.
    • The technique proved effective in experimental models of nephritis.

    Conclusions:

    • Combining TUNEL staining with cell-specific markers on isolated glomeruli is a powerful technique for studying glomerular cell turnover.
    • This enhanced method offers improved sensitivity for detecting apoptotic cells in kidney research.
    • The approach holds potential for advancing the understanding and diagnosis of kidney diseases.