Flow Cytometry Core -

A. Calibur Startup

Check the sheath fluid by lifting the 2 tanks (sheath fluid on the left and waste on the right). If the clean sheath fluid card box container is empty, replace it with a new card box sheath fluid container. Keep the empty card box, add 2 liters of pure bleach (under the sink) and replace the waste container. Please write WASTE on it and indicate the date.
Turn ON the fluidic pump (up).
Shutdown the computer if on.
Turn on the Calibur (bottom right side). Wait 1 minute.
Turn on the Computer.
Flush the fluid system of the Calibur 1-2 times by hitting the PRIME button. (Note: Pushing the PRIME button will make it will turn red. Wait till the instrument goes back into STANDBY mode before priming again.)
Log in (ID=first name initial + last name and password=first name).
Open Cell Quest
- Acquire/Connect to cytometer (browser will come up)
- Open an existing cell quest document or draw the FACS plot on new document (Make sure when drawing a FACS plot that “acquisition” is selected in the Inspector window that will pop up)
- Select Cytometer/DetectorAmp
- If previous experiments with similar instrument setting, select Cytometer/Instrument Settings:
- Open an instrument setting file or an fcs file of previous samples
- Select Set
- Select Done
- In Browser window, change the directory (the place where your file will be stored)
- Select Acquire/Acquisition and Storage, select the number of cells that to be acquired
Run your control in a Setup mode to unsure all voltages are OK.
Deselect the Setup option and hit acquire when ready to acquire controls and samples.
Copy your file on the Gladstone servers or on Vishnu
- In finder menu, select go/connect to server
- Enter the address (vishnu.ucsf.edu)
- Select FlowcoreDataA
- Copy your folders in your own Folder
Log out.

B. Calibur Shutdown

Run 10% bleach on HIGH for 5 minutes, then deionized water on HIGH for 3 minutes.
Leave the deionized water on the Calibur and turn off the machine.
Be sure to log out of the Cell Quest software.
Switch off the Calibur if needed (i.e. if you did not talk to the next user).