Instruction Manual

46 VR NxT| Instruction Manual 1.0 02-2019|VRNXTinstr1EN|© 2019 Menarini Silicon Biosystems SpA
ANNEX A: CELLSEARCH sample preparation for DEPArray
Open the CELLTRACKS cartridge
1. Hold the CELLTRACKS cartridge in one hand.
2. Insert the wide-end of a clean P1000 tip into the opening in the backside of the cartridge, against the top of the
insert.
3. Gently push the top edge of the insert to remove the lid of the cartridge.
Extract the sample from the CELLTRACKS cartridge
1. Take the P200 pipette with the BSA coated tip and push the pipette knob to the first stop.
2. Insert the tip in the CELLTRACKS cartridge and slowly release the pipette knob to aspirate the sample.
3. Slowly pipette up and down five times to suspend the sample before the final aspiration.
4. Transfer 200 μl of sample into the Sample tube.
5. Repeat steps 3 and 4 to collect any sample residual and transfer it to the Sample tube.
6. Eject the tip in the Tip tube for later use.
Wash the CELLTRACKS cartridge
1. Attach a new clean gel loading tip to the P200 pipette.
2. Aspirate 200 μl of DABUF from a DABUF tube.
3. Place the tip in the upper part of the CELLTRACKS cartridge and slowly push the pipette knob to release the
DABUF.
4. Repeat steps 2 and 3 for the remaining DABUF (~125 μl).
5. Discard the tip.
6. Take the BSA coated tip from the Tip tube and attach it to the P200 pipette.
7. Push the pipette knob until the first stop so that it is ready for aspiration.
8. Insert the pipette tip into the CELLTRACKS cartridge and gently pipette up and down five times to wash the inner
surface of the cartridge.
9. Transfer the entire sample into the Sample tube.
NOTICE: Do not discard the tip.
10. Repeat steps from 2 to 9 using the second DABUF tube.
NOTICE: Do not discard the CELLTRACKS cartridge.
11. Discard the tip.
12. Centrifuge the Sample tube at 1,000 g for 5 minutes at room temperature in a swing bucket centrifuge.
13. Carefully remove the Sample tube from the centrifuge and move it under the biosafety cabinet class II.
Wash the sample for the first time
1. Take a P1000 pipette and attach a clean P1000 tip to it.
2. Push the pipette knob so that it is ready for aspiration.
3. Without disturbing the cell pellet, remove the supernatant from the Sample tube until leaving ~50 μl of liquid.
4. Transfer the supernatant in the S1 tube.
5. Discard the tip.
6. Attach a new clean P1000 tip to the P1000 pipette.