A SpinVessel® customer conducted a feasibility study using our 850mL SpinVessel®, VP 418SV3-1-850RB. They used paramagnetic polystyrene microparticles. Aliquots were taken using a standard 1000ul pipette. The summary and results below are directly from the customer.
Feasibility Testing of the 850mL Spin Vessel
The 850mL larger-scale V&P baffled SpinVessel® was assessed for its ability to maintain a homogeneous bead solution. 8.55mL of previously coupled CTDE beads at 7500 bd/uL and 1.725mL of beads at 15000 bd/uL were combined and diluted into a 900mL BSA/trehalose solution. The beads and solution were mixed together in a 1L polystyrene bottle and rolled/mixed for 30 minutes. Upon mixing, 1mL of the bead solution was diluted into 9 mL of Isoton diluent and counted on the Coulter Counter as a control.
850mL of the diluted and mixed bead solution was then placed into the 850mL baffled SpinVessel® and placed on the V&P Stirrer. Settings of speed at 100, rotate to 5, and pause to 5 were selected for this testing.
While the bead solution was mixed on the V&P SpinVessel® mixer, 1mL of the solution was sampled at 5-minute intervals. While each 1mL sample was diluted into 9mL of Isoton and counted, another 100mL of the solution was set aside after each sampling to replicate depletion of the bulk overfilling.
The results of the Coulter Counter readings were as follows
Time Point | Sample Time | Bulk Vol (mL) | Read 1 (bd/mL) | Read 2 (bd/mL) | Avg Read | % Recovery |
---|---|---|---|---|---|---|
0(control) | 00:00 | 900 | 79000 | 82320 | 80660 | 100% |
1 | 00:05 | 850 | 78960 | 82840 | 80900 | 100% |
2 | 00:10 | 750 | 77120 | 80520 | 78820 | 98% |
3 | 00:15 | 650 | 78060 | 82540 | 80300 | 100% |
4 | 00:20 | 550 | 78360 | 78420 | 78390 | 97% |
5 | 00:25 | 450 | 77200 | 82400 | 79800 | 99% |
6 | 00:30 | 350 | 81120 | 77100 | 79110 | 98% |
7 | 00:35 | 250 | 81100 | 78820 | 79960 | 99% |
8 | 00:40 | 150 | 79720 | 81760 | 80740 | 100% |
9 | 00:45 | 50 | 81660 | 79640 | 80650 | 100% |
10 | 00:50 | 25 | 80840 | 78860 | 79850 | 99% |
11 | 00:55 | ~10 | 78920 | 80620 | 79770 | 99% |
The customer’s name and affiliation are kept confidential per request.