PHM450 cellular tox assays
Tobi Limke・17 minutes read
Viability and cytotoxicity assays are essential for measuring cell health and death, with a focus on identifying stressed or damaged cells using various techniques and markers. Specific assays like calcium dysregulation with Fura 2 a.m. and caspase activation in apoptosis kits are discussed as key methods for assessing cellular toxicity and identifying different cell death pathways.
Insights
- Viability assays focus on determining if cells are alive and metabolically active, while cytotoxicity assays measure cell death caused by toxic substances, providing insights into cell health and life status.
- Different assays, such as monitoring calcium dysregulation with fluorescent dyes like Fura 2 a.m. or assessing redox status with glutathione kits, offer valuable insights into cellular vulnerabilities and responses to stressors, showcasing the diverse approaches available to study cellular toxicity.
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Recent questions
What do viability assays determine?
Cell health and metabolic activity.
How are cytotoxicity assays different from viability assays?
They measure cell death due to toxic insult.
What is the significance of calcium dysregulation in cells?
It can lead to cell death.
How do glutathione assays help in monitoring cell vulnerability?
By indicating redox status.
What is the role of caspase-3 in apoptosis kits?
It serves as a convergence point for initiators.
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Summary
00:00
Cellular Toxicity Assays: Viability and Cytotoxicity Measurements
- Lecture covers practical assays for measuring cellular toxicity
- Focus on recognizing basic in vitro assays and linking them to endpoints
- Assays aim to identify stressed, damaged, or dead cells
- Viability assays determine if cells are living and metabolically active
- Cytotoxicity assays measure cell death due to toxic insult
- Viability assays focus on cell health and life status, while cytotoxicity assays count living and dead cells
- Assays can observe single cells or be automated for high-throughput screening
- Calcium dysregulation can lead to cell death, measured using fluorescent dyes like Fura 2 a.m.
- Glutathione assays monitor redox status, indicating cell vulnerability to oxidative stress
- Glutathione kits are relatively inexpensive and require a spectrophotometer for reading absorbance
19:39
Assays for Apoptosis and Necrosis Detection
- Caspase-3 is a key focus in apoptosis kits due to its early activation and role as a convergence point for multiple initiators, but these assays only measure apoptosis, not necrosis, as necrosis does not involve caspase activation.
- Specific examples of assays are discussed, such as live and dead cell staining using a combination of thid 'i'm homodimer and calcium am, measuring ATP levels with luciferase, and using stauros pouring as a positive control in caspase-3 assays.
- Fura 2 a.m. is utilized to measure intracellular calcium concentrations, while trypan blue exclusion assay distinguishes between live and dead cells based on membrane integrity, with dead cells taking up the dye.
