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Protocol to study cell death using TUNEL assay in Drosophila imaginal discs
Anuradha Venkatakrishnan Chimata1, Prajakta Deshpande1, Abijeet Singh Mehta1
1Department of Biology, University of Dayton, Dayton, OH 45469, USA.
STAR Protocols
|February 7, 2022
Summary
The TUNEL assay effectively labels and quantifies cell death by detecting DNA fragmentation in Drosophila eye-antennal imaginal discs, aiding research into diseases linked to cell death misregulation.
Area of Science:
- Developmental Biology
- Cell Biology
- Genetics
Background:
- Cell death is crucial for maintaining tissue homeostasis.
- Dysregulation of cell death contributes to diseases such as cancer and neurodegeneration.
- The Terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay is a key tool for detecting DNA fragmentation during cell death.
Purpose of the Study:
- To utilize the TUNEL assay for labeling and quantifying cell death.
- To apply this method in Drosophila melanogaster third-instar larval eye-antennal imaginal discs.
- To assess the sensitivity of the TUNEL assay in detecting apoptosis in retinal neurons.
Main Methods:
- Application of the TUNEL assay on Drosophila melanogaster larval eye-antennal imaginal discs.
- Fluorescent labeling of fragmented DNA in dying cells.
- Microscopic detection and quantification of labeled cells.
Main Results:
- The TUNEL assay successfully labeled fragmented DNA in dying cells within the Drosophila eye-antennal imaginal discs.
- Quantification of cell death was achieved using this method.
- The assay demonstrated sensitivity in detecting DNA fragmentation during apoptosis in retinal neurons.
Conclusions:
- The TUNEL assay is a reliable method for studying cell death in Drosophila eye-antennal imaginal discs.
- This technique aids in understanding the role of apoptosis in developmental processes and disease.
- Further research can leverage this assay to investigate cell death mechanisms in various biological contexts.