What is PI staining?
Description. Propidium iodide (PI) is a popular red-fluorescent nuclear and chromosome counterstain. Since propidium iodide is not permeant to live cells, it is also commonly used to detect dead cells in a population. PI binds to DNA by intercalating between the bases with little or no sequence preference.
Does PI stain apoptotic cells?
The ability of PI to enter a cell is dependent upon the permeability of the membrane; PI does not stain live or early apoptotic cells due to the presence of an intact plasma membrane 1,2,6.
Is Pi toxic to cells?
Furthermore, 0.1 μM PI was found to be non-toxic and universally applicable, as revealed by testing a wide range of microorganisms cultivated in different complex media, including Micrococcus luteus (1.78% PI+), Bacillus subtilis (0.09% PI+), E. coli (<0.01% PI+), Vibrio harveyi (<0.01% PI+) and the yeast S.
Where does rhodamine 123 stain in the cell?
mitochondria
Rhodamine 123 is a popular green fluorescent mitochondrial dye that stains mitochondria in living cells in a membrane potential-dependent fashion. It is widely used in flow cytometry studies involving mitochondrial membrane potential.
How do you dissolve pi?
To make a stock solution from the solid form, dissolve PI (MW = 668.4) in deionized water (dH2O) at 1 mg/mL (1.5 mM) and store at 4°C, protected from light. When handled properly, solutions are stable for at least 6 months.
How to stain a fixed cell with Pi?
Staining cells with PI Centrifuge the ethanol fixed cells 5 min at 300 x g and decant ethanol thoroughly (be careful not to lose your cells!). Suspend the cell pellet in 5 ml PBS, wait 60 sec, and centrifuge 5 min at 200 x g. Suspend the cell pellet in 1 ml PI staining solution that has been optimized for your cell type and concentration.
How to stain propidium iodide ( PI ) in PBS?
Resuspend the cells in 0.7 ml PBS, vigorously vortex the pellet for 10 seconds, and add 10 ml 75% ethanol – vortex to mix briefly. Fix cells at 4 o C O/N or until ready to stain. Briefly vortex then spin fixed cells down at 3000 rpm for 5 min. Pour off ethanol. Leave less than 0.2 ml ethanol in tube.
How does ethanol affect the staining of Pi?
For optimal PI staining, cells require permeabilization with ethanol to allow the dye access to the nucleus. However, such treatment with ethanol results in the loss of GFP fluorescence as the soluble, cytoplasmic GFP leaks out of the cells following permeabilization.
What kind of staining is used for propidium iodide?
Propidium Iodide (PI) Staining. Sorting and Staining Buffers. 2% Paraformaldehyde. **These protocols are for staining the DNA of all cells in a sample for use in DNA cell cycle analysis . They are not recommended for apoptosis analysis, except as a preliminary check for DNA degradation (subG0).