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Identifying and quantifying apoptosis: navigating technical pitfalls.

Megan M Garrity1, Lawrence J Burgart, Darren L Riehle

  • 1Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, Minnesota 55905, USA. garrity.megan@mayo.edu

Modern Pathology : an Official Journal of the United States and Canadian Academy of Pathology, Inc
|April 15, 2003
PubMed
Summary
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This study optimized the TUNEL method for apoptosis detection using BLISS imaging. The enhanced technique improves accuracy and reduces false positives in malignancy research.

Area of Science:

  • Oncology
  • Cell Biology
  • Biotechnology

Background:

  • Apoptosis is crucial in cancer, often assessed by TUNEL.
  • Standard TUNEL kits can yield high background and false positives, complicating apoptosis vs. necrosis distinction.

Purpose of the Study:

  • To develop a rapid, reproducible method for detecting and quantifying apoptosis.
  • To overcome limitations of standard TUNEL protocols using advanced imaging.

Main Methods:

  • Optimized the Apoptag Plus Peroxidase In Situ Apoptosis Detection kit.
  • Integrated quantitative histomorphometric computer imaging with the Bacus Laboratories Incorporated Slide Scanner (BLISS).
  • Scanned multiple high-resolution images, analyzed using WebSlide Browser, considering immunohistochemistry and surrounding cell histology.

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Main Results:

  • The BLISS imaging system significantly reduced false positive and negative staining in apoptosis detection.
  • Simultaneous review of equivocal staining was facilitated via networked access.
  • Quantitative analysis incorporating histology improved accuracy over TUNEL alone.

Conclusions:

  • Optimized BLISS imaging offers a superior method for apoptosis quantification in malignancies.
  • This approach mitigates common drawbacks associated with the TUNEL technique.
  • The method enhances diagnostic accuracy and reproducibility in cancer research.