BD LSRFortessa User manual

flowkon
Flow Cytometry User Instructions
at the University of Konstanz
FACSFortessaTM –HTS User Instructions
The BD™ High Throughput Sampler (HTS) option for the BD LSRFortessa™ cell analyzer
provides rapid, fully automated sample acquisition from microtiter plates. In high-
throughput mode, the HTS can process a 96-well plate in fewer than 15 minutes.
Standard throughput mode can be selected for acquisition of larger sample volumes.
Preparing the HTS
Switch the acquisition control to plate mode.
This will switch off the DCM pump.
Unscrew the tube retainer (grey ring ) that holds the
DCM sleeve onto the SIT and carefully remove the sleeve.
Remove the DCM sleeve and replace it with the shorter SIT
protector.
Install the sample coupler on the SIT.
Check the level in the connected water bottle (> ~30 mL).
Switch on the HTS reader (on the backside).
Press the Run button (the HTS does not operate in STNDBY mode).
Initialisation and Priming the HTS
Note: Don’t change PMTV of fluorescence channels after enabling compensation.
Operation of the loader is only possible via DIVA software –do not use the prime
button on the instrument.
When the BD DIVA software is already open, go HTS Re-Initialize.
Start BD DIVA software. The HTS will initialize automatically, otherwise go to HTS
Re-Initialize
Prime the HTS (Sheath tubing connected): HTS Prime. 3x in total.
Run your Experiment
See detailed instructions on HTS Programming.

Cleaning Procedure after Measurement
Include 3x4 wells for cleaning solutions on your plate (F9-F12, G9-G12, H9-H12)
OR use a separate plate for the cleaning procedure.
Fill 250 µL FACSClean in 4 wells, F9-F12.
Fill 250 µL FACSRinse in 4 wells, G9-G12.
Fill 250 µL MQ H2O in 4 wells, H9-H12.
Shutting Down the System
Return to Tube-based acquisition mode:
STNDBY mode.
Switch off HTS.
Remove the sample coupler from the SIT and re-install the standard DCM sleeve.
Switch to Tube mode.
Prime 3x with arm at side.
Install MQ-water tube, close aspirator arm.
Close open experiments, close the DIVA software, turn off instrument, fluidic cart
and PC.
Provide information about your measurement in the LogBook.