Table 2. Sorting efficiency with larger number of cells. We used a mixed culture of unlabeled NE-4C and fluorescent NE-GFP-4C cells in 6 experiments with a total number of 311 cells selected for sorting. After a brief (30s) trypsin-EDTA treatment software detected the cells in a merged image of the culture captured in both phase-contrast and fluorescent modes of the microscope. Vacuum was in the range of [−9,000; −11,000] Pa in the syringe connected to the micropipette with an aperture of 70 μm. After the software excluded those cells that were closer to other cells than 70 μm we picked up fluorescent cells automatically. Cells were then injected to a 6 mm PDMS cylinder attached onto a glass cover slip to make easier the subsequent search for cells. 2 h later we counted live fluorescent and unlabeled cells on the cover slip. Pick up efficiency was calculated as the ratio of the number of selected cells picked up and the total number of selected cells. Sorting purity is the ratio of the number of fluorescent cells and the total number of cells picked up. Average sorting purity and pick up efficiency was 95±2% and 93±5%, respectively. In some experiments the micropipette picked up extra fluorescent cells from the neighborhood of cells selected for sorting. The number of these cells is indicated after the + signal in the 'Number of fluorescent cells picked up' column. The rate of survival was calculated as the ratio of the number of cells survived and the number of cells picked up including also unselected cells. Average rate of survival was 62±7%. Average purity of the sorted culture (number of fluorescent cells divided by the total number of cells) was 95±2%. Average rates were calculated as the sample mean weighted by the denominator of each ratio, i.e., the number of input cells. The error of the mean was approximated by the weighted sample variance divided by the square root of the number of experiments.

Number of cells selected for sorting	Number of fluorescent cells picked up	Number of unlabeled cells picked up	Sorting purity	Pick up efficiency	Number of fluorescent cells survived	Number of unlabeled cells survived	Rate of survival	Purity of the sorted culture
86	85+2	1	0.99	0.99	40	0	0.45	1
53	38	2	0.95	0.72	16	1	0.42	0.94
65	59	10	0.86	0.91	49	7	0.83	0.88
19	19	1	0.95	1	14	1	0.74	0.93
42	42	0	1	1	25	0	0.6	1
46	46+1	1	0.98	1	37	1	0.79	0.97