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Mechanism of Regulation of Adipocyte Numbers in Adult Organisms Through Differentiation and Apoptosis Homeostasis
Published on: June 3, 2016
Detection of apoptosis in tissue sections.
Eva Csizmadia1, Vilmos Csizmadia
1Center for Vascular Biology, Beth Israel Deaconess Medical Center, Boston, MA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|July 18, 2009
Summary
This study details a reliable protocol for detecting apoptotic cells using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assays in animal disease models. The method also allows for simultaneous protein immunodetection in tissues.
Area of Science:
- Cell Biology
- Histology
- Pathology
Background:
- Apoptosis is a critical cellular process implicated in various diseases.
- Accurate detection of apoptotic cells in tissue is essential for disease research.
- Standard histological staining methods can be challenging for apoptotic cell identification.
Purpose of the Study:
- To present a refined protocol for TUNEL-based detection of apoptotic cells in animal tissues.
- To optimize tissue preparation and fixation for enhanced histological staining.
- To enable simultaneous immunodetection of proteins alongside apoptosis assays.
Main Methods:
- Utilized TUNEL (terminal deoxynucleotidyl transferase dUTP nick end labeling) assays.
- Emphasized meticulous tissue preparation and fixation techniques.
- Employed DAB (3,3'-diaminobenzidine) or fluorescence-based immunostaining for protein detection.
Main Results:
- Successfully demonstrated the presence of apoptotic cells in target tissues of animal disease models.
- Achieved reliable detection of TUNEL-positive cells.
- Validated the co-detection of specific proteins within apoptotic and surrounding cells.
Conclusions:
- The presented protocol ensures reliable TUNEL-based apoptosis detection in diverse animal disease models.
- Optimized tissue handling is critical for successful histological and immunofluorescence analyses.
- This method facilitates comprehensive cellular and molecular analysis of diseased tissues.
