To begin, switch on the optics based measurement system, microscope light emitting diode source, and the computer. Start the program on the computer and open a new file by clicking new under the file in the top left of the screen. Click on collect, choose experiments and then select IPSC and calcium measurements.
Next, replace the human IPSC cardiomyocyte culture medium with 0.5 milliliters per well of Tyrode's solution. In a 24 well plate. Replace the plate lid with the climate control lid and place the plate inside the optics based measurement system in the device adjusted to 37 degrees Celsius temperature.
Click on open cell finder under the toolbar and wait for a new screen to appear. Select the plate format under chamber type and well at the top of the screen. Select a well and choose an area within the well to observe synchronized contraction and relaxation.
Press start on the bottom left of the screen. Then click add measurement to measure the area. Measure approximately 10 areas per well in a 24 well plate.
For only baseline measurements, continue the measurement from the next well. For testing the compounds, press complete once the areas within a well are measured. Open the optics based measurement instrument and add the desired stressor or compound to the well.
After incubating at the desired time period, click on automatic measurement to perform paired measurements in the same areas of the well where the baseline measurements were performed. Press complete after all the measurements are performed. Choose the next well from the top right of the screen and continue measurements for all the required wells.
After measuring all the required wells, click stop and save the file to the desired location.
