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Q4: How long can cells be stretched?
A4: The duration depends on cell strain and condition. In general, two weeks
cultivation in the incubator is possible with culture media changes every 2-3 days. It
is also important to monitor reservoir coolant for the instrument motor. Insufficient
amount of the cooling water may increase the temperature of the motor resulting in
killing of cells and /or burning out the motor.
Q5: How can I obtain protein or mRNA samples from the cells attached to the silicone
membrane?
A5: (1) Proteins for Western blotting: Wash the cels once with PBS. Add SDS-
PAGE sample loading dye directly into the chamber, and collect the cell extract by
using a cell scraper.
(2)Proteins for Immunoprecipitation: Wash the cells once with PBS. Add cell extract
buffer directly into the chamber, and collect the cell extract by using a cell scraper.
(3)RNA: Wash the cells once with PBS (for RNA preparation). Add RNA extraction
buffer directly into the chamber, and collect the cell extract by using a cell scraper.
Q6: I want to use recombinant cells for an experiment.
A6: Direct transfection of cells in the strain chamber may be possible. However,
transfection itself may damage the cells, which may make getting clear image data
difficult. We recommend performing the transfection in a normal culture dish then
transferring the recombinant cells into the strain chamber.
Q7: Cells seem to be crowded in the center of the chamber instead of being uniformly
distributed throughout the chamber.
A7: Vibration from the incubator may disrupt the distribution of the cells. We
recommend gently rocking the chamber 15 minutes after seeding your cells.
