- Display and control panel
- Waste reservoir
- Buffer reservoir
- Sample vacutainer
- Cassette clamp
- Harvesting valve
- Reagent lines and tubes:
Before you start, check…
Blood sample preserved in EDTA, less than 48 hours old, stored at 4°C
An empty vacutainer and a cleaning cassette in place.
Reagent tubes 1-6 are empty and the lines are clean.
The harvesting valve is turned clockwise.
Reagent volumes are topped up:
- 100 ml PBS in buffer reservoir
- 50 ml Distel in waste reservoir (note: empty waste when it reaches 400 ml and replace Distel)
- 20 ml Decon Decomatic (10%) in cleaning solution tube (label C)
- 20 ml ethanol (100%) in priming solution tube (label P)
- Select protocol PX2_P and press Run then Start.
- On prompt remove the cleaning cassette and insert a new cassette.
- Press OK to start the priming cycle.
- When the cycle is finished (after 15 min), press OK then Continue.
- Select protocol PX2_S99F and press Run then Start.
- At the “Rinse vacutainer” prompt, partially remove the vacutainer off its mount keeping the line inside the tube.
- Press OK to start the rinse and collect the fluid in the vacutainer. Using an ethanol soaked tissue, wipe the outside of the line and the O-ring area at the top of the mount. If needed, clean the O-ring with 1% Distel before wiping with ethanol.
- On prompt, attach the sample vacutainer and press OK to start the separation cycle.
- When the cycle is finished (after 2.0 h for a 10 ml sample), press OK then Continue.
- After a middle clean cycle, remove the cleaning cassette and insert the separation cassette.
- Select the protocol PX2_H and press Run then Start.
- On prompt, rotate the harvesting valve anticlockwise and press OK.
- Remove the harvest line from the waste collection tube and wipe with an ethanol soaked tissue. Prepare the collection vessel (e.g. Eppendorf tube).
- Press OK to start the harvest. A volume of 200 l will flow through the harvest line. If required, press YES to collect a further 1 ml flush.
- On prompt, rotate the harvest valve clockwise and press OK. When finished (after 5 min) press OK then Continue.
Note: Please reference ‘Parsortix in-cassette staining’ guide for fuller instruction
- Add the following reagents to clean empty 15 ml Falcon tubes:
- Line 1: 2 ml 4% formaldehyde
- Line 2: 2 ml 0.1% Triton X-100 in PBS
- Line 3: 1 ml blocking buffer
- Line 4: 1 ml primary antibodies (4 l each antibody) in blocking buffer
- Line 5: 1 ml secondary antibodies (1 l each antibody and 0.2 l DPI) in blocking buffer
- Select protocol PX2_stain4h and press Run then Start.
- When finished (after 6.5 h) press OK then Continue.
- Select the appropriate cleaning protocol according to the flowchart and press Run then Start.
- Middle clean: PX2_CT2, before cell harvest, 25 min
- Short clean: PX2_CT, after cell harvest, 45 min
- Full clean: PX2_C, after in-cassette staining, 1 h 15 min
- On prompt, remove the separation cassette, insert a cleaning cassette and press OK.
- Empty the reagent tubes if necessary and press OK to start the cleaning cycle.
- When finished press OK then Continue. After a full clean cycle, empty the reagent tubes 1-6 and the harvest tube. Wipe the outside of the lines if they have been sitting in fluid.