iv. If this does not resolve issue, check the optical filters, sheath
tank seal, check front cytometer pane for liquid levels.
3. If you fill the di water bottle you will need to de-bubble the di
water filter using the option in the software.
4. Sorting Setup
1. ALL SAMPLES, INCLUDING COMPS, MUST BE
2. Select the experiment you would like to use, or the SSFF
3. On the upper right, change the experiment name.
4. Fill in other information for your experiment.
5. Check or uncheck the fluorescent parameters that are
needed, and name the parameters if you wish.
6. Turn on/off lasers for your experiment. Be sure that the
488nm laser is on. The violet laser decreases the life of
the chip. Only turn it on if needed.
7. Click Create New Experiment in the lower right of the
When running a compensation matrix
1. There are 2 methods for compensation available the wizard and manual
2. Choose to start compensation wizard if you are sorting with multiple
fluorochromes and have prepared single color controls.
3. Click OK to continue
4. Follow the wizard's instructions and run the negative control, and
adjust the instrument settings in detector/threshold settings FSC
and BSC to get population on scale. Briefly load a fully stained
sample at this point (do not record it) and verify that none of the
positive populations are off scale. If they are, lower the PMT
value to get them back on scale.
5. Then record negative and adjust the gate such that it is around the
6. Positive sample using these adjusted settings.
7. Acquire single-stained controls, then set compensation as directed.
8. Click Close then Finish to end the Wizard.