What initiates cell death?

What initiates cell death?

Apoptosis is mediated by proteolytic enzymes called caspases, which trigger cell death by cleaving specific proteins in the cytoplasm and nucleus. Caspases exist in all cells as inactive precursors, or procaspases, which are usually activated by cleavage by other caspases, producing a proteolytic caspase cascade.

How do you detect cell death?

Necrosis is detected by measuring the permeability of the plasma membrane to a normally impermeable fluorescent dye, such as the DNA-binding dye propidium iodide (PI). Apoptosis is detected by measuring the externalization of phosphatidylserine on the plasma membrane using fluorescent-tagged annexin V.

Does Hoechst require Permeabilization?

Hoechst dyes are cell-permeable so there is no need to permeabilize them for Hoechst staining.

What are the 3 stages of apoptosis?

apoptosis

• Induction phase.
• Early phase.
• Mid phase.
• Late phase.

Which processes are part of the initiation phase of apoptosis?

The apoptotic death pathway can be initiated by a variety of stimuli, including DNA damage, the withdrawal of growth factors, and the binding of certain ligands to cell surface receptors.

Which initiates cell division in animal cell?

centriole
The cell organelle responsible for initiating cell division is the centriole. It produces mitotic spindle fibres which are a crucial part of the cell division. The centriole is also involved in cytokinesis, where the cytoplasm begins to divide, resulting in two daughter cells.

What is cell death assay?

Cell viability and cytotoxicity assays measure cellular or metabolic changes associated with viable or nonviable cells. These assays can detect structural changes such as loss of membrane integrity upon cell death or physiological and biochemical activities indicative of living cells.

Which is better to stain dead cells with DAPI or Hoechst?

In S. cerevisiae, DAPI and Hoechst preferentially stain dead cells with nuclear and cytoplasmic localization. In live yeast, Hoechst shows dim nuclear and cytoplasmic staining, while DAPI shows dim mitochondrial staining.

Which is better to stain live cells or dead cells?

Dead cells tend to stain more brightly than live cells. In S. cerevisiae, DAPI and Hoechst preferentially stain dead cells with nuclear and cytoplasmic localization. In live yeast, Hoechst shows dim nuclear and cytoplasmic staining, while DAPI shows dim mitochondrial staining. The dyes can be used to stain yeast at 12-15 ug/mL in PBS.

How much Hoechst can be used to stain cells?

Dilute Hoechst dyes to 10 ug/mL, or dilute DAPI to 100 ug/mL. Note: DAPI or Hoechst can be combined with other fluorescent probes. Without removing the medium from the cells, add 1/10 volume of 10X dye directly to the well.

How long to incubate cells for nuclear staining?

For larger well sizes (e.g., 24-well to 6-well plates), the plate can be gently swirled to mix. Incubate cells at room temperature or 37°C for 5-15 minutes, then image. Note: Washing is not necessary for specific staining, but nuclear staining is stable after washing.