Detection of Changes in Mitochondrial Membrane Potential in T-cells Using JC-1 Dye: Version 1
- PMID: 39013043
- Bookshelf ID: NBK604957
- DOI: 10.17917/RTMF-4853
Detection of Changes in Mitochondrial Membrane Potential in T-cells Using JC-1 Dye: Version 1
Excerpt
The purpose of this protocol is to understand the propensity of nanomaterials to induce mitochondrial damage as assessed by change in mitochondrial membrane potential (ΔФm). Alteration in mitochondrial physiology is commonly linked with apoptotic cell death. Under stressed conditions, the mitochondrial permeability transition pores open, leading to a decrease in mitochondrial membrane potential [1]. The changes in ΔФm further activates various signaling cascades leading to cytochrome C release, ATP depletion and accumulation of reactive oxygen species (ROS), leading to cell death via either necrosis or apoptosis [1, 2]. The combination of mitochondrial membrane potential loss and mitochondrial ROS lead to organelle specific toxicity that ultimately cause cell death.
Sections
- 1. Introduction
- 2. Principles
- 3. Reagents, Materials, and Equipment
- 4. Reagent and Control Preparation
- 5. Preparation of Nanoparticles
- 6. Collection and Handling of Whole Blood
- 7. Preparation of T-Cells
- 8. Treatment with Nanoparticles and Controls
- 9. Flow Cytometry
- 10. Data Analysis and Report
- 11. Acceptance criteria
- 12. References
- 13. Abbreviations
- 14. Appendix
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References
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- Hirsch, T., Marzo I., and Kroemer G., Role of the mitochondrial permeability transition pore in apoptosis. Biosci Rep, 1997. 17(1): p. 67–76. - PubMed
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