ANGLE’s Parsortix system has the potential to deliver profound improvements in clinical and health economic outcomes in the treatment and diagnosis of various forms of cancer.”
Andrew Newland
Chief Executive
Parsortix™ PR1 (For Research Use Only)

System Components

  1. Display and control panel
  2. Waste reservoir
  3. Buffer reservoir
  4. Sample vacutainer
  5. Cassette clamp
  6. Harvesting valve
  7. Reagent lines and tubes:
Parsorter_PR1

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)

Parsortix Workflow

Parsortix_UK_Workflow_Chart

Priming

  1. Select protocol PX2_P and press Run then Start.
  2. On prompt remove the cleaning cassette and insert a new cassette.
  3. Press OK to start the priming cycle.
  4. When the cycle is finished (after 15 min), press OK then Continue.

Blood Separation

  1. Select protocol PX2_S99F and press Run then Start.
  2. At the “Rinse vacutainer” prompt, partially remove the vacutainer off its mount keeping the line inside the tube.
  3. 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.
  4. On prompt, attach the sample vacutainer and press OK to start the separation cycle.
  5. When the cycle is finished (after 2.0 h for a 10 ml sample), press OK then Continue.

Cell Harvest

  1. After a middle clean cycle, remove the cleaning cassette and insert the separation cassette.
  2. Select the protocol PX2_H and press Run then Start.
  3. On prompt, rotate the harvesting valve anticlockwise and press OK.
  4. Remove the harvest line from the waste collection tube and wipe with an ethanol soaked tissue. Prepare the collection vessel (e.g. Eppendorf tube).
  5. 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.
  6. On prompt, rotate the harvest valve clockwise and press OK. When finished (after 5 min) press OK then Continue.

In-cassette staining

Note: Please reference ‘Parsortix in-cassette staining’ guide for fuller instruction

  1. 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
  2. Select protocol PX2_stain4h and press Run then Start.
  3. When finished (after 6.5 h) press OK then Continue.

Cleaning

  1. Select the appropriate cleaning protocol according to the flowchart and press Run then Start.
    • Pre-Harvest Flush: PX2_CT2, to be used ONLY 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
  2. On prompt, remove the separation cassette, insert a cleaning cassette and press OK.
  3. Empty the reagent tubes if necessary and press OK to start the cleaning cycle.
  4. 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.
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