Cellstain-Double Staining Kit

Protocol tips

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	Mix 2μM calcein AM and and 4 μM propidium iodide and incubate the mixture for 15 minutes at 37C

Protocol tips
Mix 2μM calcein AM and and 4 μM propidium iodide and incubate the mixture for 15 minutes at 37C

Publication protocol

Live/dead assay was performed with calcein-AM (Dojindo, Kumamoto, Japan) and propidium iodide (Sigma-Aldrich, St. Louis, MO). A solution containing 2 μM calcein AM (green- fluorescent) to indicate intracellular esterase activity, and 4 μM propidium iodide (red-fluorescent) to indicate loss of plasma membrane integrity was introduced into the cell- patterned channel, and incubated for 30 min at 37°C and 5% CO2.

NucView 488 caspase-3 assay kit (Biotium, CA, USA) wasused for the apoptosis assay. A solution containing 5 μM NucView 488 caspase-3 substrate in binding buffer was placed on a glass substrate with cells, and incubated for 30 min at 37°C and 5% CO2. The nuclei of apoptotic cells were stained green. Cells were visualized using a fluorescence microscope IX71 with a dichroic mirror block (U-MWIBA3 and U-MWIG3 Olympus; for cells stained with green and orange or red dye, respectively).


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