Kits include a premixed solution of AO and PI in PBS. The reagents should be stored protected from light at 2 – 8oC in an airtight container.
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- Equilibrate all solutions to room temperature.
- Vortex cell suspension and AO/PI solution prior to use.
- For each sample, mix AO/PI and a cell suspension together in a 50% solution.
- Note: A 50% solution is made up of one part AO/PI and one part cell suspension, resulting in a Dilution Factor of 2 for the cell suspension.
- Mix sample thoroughly prior to loading onto the CellDrop.
- Ensure that the arm is in the down position, and launch the AO/PI app.
- Clean the sample surfaces if there is visible debris in the preview image.
- Optional: Enter a sample name and any additional sample information.
- Select or create a protocol, and set the Dilution Factor to 2 for a 1-to-1 dye/cell suspension mix.
- With the arm down, dispense the sample aliquot into the measurement chamber using the groove on the lower sample surface as a pipetting guide.
- Note: The volume of the sample required depends on the protocol settings for the chamber height. The required volume is displayed on the Count button.
- Use the brightfield channel to adjust the focus. Verify focus in green channel prior to count.
- Switch to the green and red channels and adjust exposure.
- When the cells have settled and are no longer moving, tap the Count button.