ii. For the 130
o You must have a filled sheath tank
o Stream appears very unstable with low sheath fluid volumes. Instability has been noted when tank is less than
50% full so make sure to Top it 0ff !
o De-bubble the filter
4. Turn pressure on before continuing on to any other steps.
5. Power up the SH800. The display screen will show the active levels of fluids.
6. Disconnect the airline in from the sheath tank. Release pressure using pressure relief ring. Fill sheath tank to below
the upper weld line. The pressure release ring on top of the tank should not be left pointing up, place it in the groove
on the top of relief valve.
7. Empty the waste tank at the sink, wear the face shield while emptying. Add bleach to the tank to the bleach line
indicated on the bottom of the carboy before replacing.
8. *Once the system has been setup, you cannot empty the waste or fill sheath without putting the machine in
"standby" mode. Doing so without putting the machine in standby mode will turn the stream off and the chip will
have to be re-calibrated. Directions for putting the machine in standby are located on page 6
9. Fill water Sterile Di tank in the left panel on the sorter with sterile water located on shelf to the right above machine
or above the hall sink shelves to the left. When the water tank has been filled, the filter must be de-bubbled through
the software. You will need to open the side access panel and vent the filter into a paper towel.
10. Log into windows using FACS User account (no password).
11. Once the SH800 has gone to standby, start the SH800 software.
12. Log in with your user name and password. If it does not ask you to scan the chip, log out and log in as "facs user."
13. Turn on aerosol management system if you are sorting a BSL-2 sample.
14. Fill out the Biohazard form on clipboard identifying the type of hazard and clean up procedure.
Automated Alignment and Sort Setup (20 minutes)
1. On log in you are prompted to scan and load the new chip – Chips are good for 24h and the previous
day's chip can be reused if <24h has passed. If continuing use on the current chip, simply open the
instrument front and reload the current chip once it is ejected. Load/ exchange the chip following the
onscreen instructions. Write date and time on the package if using a new chip. Put the old chip in the
previous day's package and put in box with used chips.
i. To load a chip – feed the chip until the soft stop then gently push into seat.
ii. Select only the
488nm blue laser
iii. Select filter configuration for
iv. Follow prompts on screen check sample line for back flush
v. Run the sheath filter de-bubble option available on the bottom of the screen during start up.
vi. Run the sample line clean if this is the first sort of the day, this runs both the bleach 12ml and DI
2. Run autocalibration–initial calibration needs only the blue laser selected.
i. When prompted, load the calibration beads. The bottle is located in the small fridge to the left of the
sink. Mix the vial gently and dispense 8 drops into a 5ml FACS tube (can be polystyrene or
polypropylene). Make sure the tube goes to the bottom of the tube holder.
ii. Step one of the calibration aligns the chip, finds the target fluorescent sensitivity this takes approximately 5
minutes. Step 2 fine tunes stream profile, side streams and drop charge.
iii. If autocalibration fails, do a sheath filter de-bubble
for initial calibration.
405nm violet laser
check box(this is our system not your application