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(Color online) Schematic illustration for separation process. (a) Loading cell suspension. (b) Turning over the coverslip. (c) Locating an inlet to inject a buffer solution and (d) a magnet. (e) Detaching the bottom of a droplet due to buffer injection. (f) Detailed cell arrangement in droplet of (d).
(Color online) Schematic drawing of imprinting process on a coverslip.
(Color online) Experimental setup. (a) Configuration of cell separation system; to effectively handle a droplet which is attached to the fabricated coverslip, we contrived a DMACS tool. A dripped cell suspension was collected by FACS tube or EP tube. (b) Working scheme for the buffer injection; this shows a process for detaching a droplet, unveiled at (c) the integrated DMACS tool.
Comparison of the separation efficiency in the MACS and the DMACS (where “positive fraction” means a relative ratio of the positive cells included in the positive fraction after cell separation and “negative fraction” means a relative ratio of the positive cell included in the negative fraction after cell separation).
(Color online) Buffer injection position (left) and separation efficiency (right).
(Color online) Demonstration of the positive cells included in each fraction: The top row, entire cells at the bright field, (a)–(e). The middle row, positive cells labeled by streptavidin-PE-Texas Red, (f)–(j). The bottom row, images merged with the top and middle rows, (k)–(o). The (+/+) means positive cells in the positive fraction as CD45 positively selected BMCs and (+/−) means positive cells in the negative fraction as CD45 depleted BMCs, (k) BMCs before separation, (l) CD45 positively selected BMCs with DMACS, (m) CD45 positively selected BMCs with MACS, (n) CD45 depleted BMCs with DMACS, and (o) CD45 depleted BMCs with MACS.
(Color online) Cell viability of the DMACS platform based on gas-liquid interface; to examine the effect of gas-liquid interface, cells obtained from the negative fraction (presented by the black-dotted line) were dyed by Hoechst 33 342, which allows viable cells to be blue color in fluorescent field. This figure shows a hanging droplet on (a) initiation and (c) completion of cell separation at bright field. Results of cell viability in each case are (b) and (d), respectively. To effectively present the results, we merged (gray-colored) entire cells at bright field with (blue-colored) viable cells at fluorescent field. The 00:00 (or 12:00) in (a) [or (c)] means the time elapsed (minute:second). The arrows in (b) and (d) indicate dead cells.
Cell loss due to adhesion and the calculated contact surface area. Percentage of : “Control” means the total number of cells “Before” separation. A contact surface means summation of the area in the separator to which cells can attach.
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