Figure 2
From: Automated single cell isolation from suspension with computer vision
Single cell isolation from a sparse suspension.
Panels (a–c) show the pick-up process of a single cell from a sparse fluorescent suspension. Arrow points to the location of the selected cell visible in (a) before picking it with the micropipette. (b) shows the same region after picking the cell. Tip of the I.D. 30 μm micropipette is visible in (b) at the location of the removed cell. (c) shows a combined image of (a,b) converted to red and green, respectively. Displacement of cells can be observed as green cells do not perfectly overlap with red ones. Green image of the selected cell is missing but all other cells remained in the dish. We compared our results to the method of single cell trapping in PDMS microwells and subsequent sorting (d,e) with a micropipette. Path of the I.D. 70 μm micropipette visiting all detected cells is shown in (d). Yellow dot marks the first cell to be picked. A significant ratio of cells (shown in (e)) adhered too strongly into the PDMS wells and thus could not be picked up. Comparison of the efficiency of the two techniques is summarized in (f). Ratio of successful single cell isolation was improved from about 50% to above 75% when using the new technique eliminating cell adhesion (Supplementary Table S1).
