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Efficient capture of circulating tumor cells with a novel immunocytochemical microfluidic device
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10.1063/1.3623748
/content/aip/journal/bmf/5/3/10.1063/1.3623748
http://aip.metastore.ingenta.com/content/aip/journal/bmf/5/3/10.1063/1.3623748
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Figures

Image of FIG. 1.
FIG. 1.

General protocol. Tumor cells (Fig. 1: 1) are incubated with biotin-tagged antibody. (Fig. 1: 2) Next, a suspension of cells is drawn through the channel. Wherever a CTC makes contact with a post, the biotin on its surface reacts with streptavidin, thus immobilizing labeled cells. (Fig. 1: 3) Next, the captured cells are fluorescently stained (Fig. 1: 4) and counted using fluorescent microscopy (Fig. 1: 5). Modifications to this procedure allow for measurement of the effects of anti-clumping reagent (Fig. 1: 6) and bystander white blood cells (Fig. 1: 7), as done in Part I. An additional pre-labeling step allows for tumor cells incubated under multiple antibody conditions to be run simultaneously in one channel at a particular flow rate, as done in Part II (Fig. 1: 8). Cell lines, antibody preparation and microchannel set-up are identical for all experiments run.

Image of FIG. 2.
FIG. 2.

Schematic of capture zones. (a) Close up of the channel.

Image of FIG. 3.
FIG. 3.

Capture efficiency vs. antigen density.

Image of FIG. 4.
FIG. 4.

Streamlines around post.

Image of FIG. 5.
FIG. 5.

k values vs. channel segment at various flow rates, plotted for four different effective antigen densities: (a) 60 000, (b) 30 000, (c) 15 000, and (d) 6000.

Image of FIG. 6.
FIG. 6.

k values vs. flow rate of various channel segments, plotted for four different effective antigen densities: (a) 60 000 (b) 30 000 (c) 15 000, and (d) 6000.

Image of FIG. 7.
FIG. 7.

Impact of flow rates upon percent of cells captured in: (a) K562 cells, (b) SKOV cells, and (c) T24cells.

Image of FIG. 8.
FIG. 8.

Effect of high flow rate on percentage of SKOV and SKBR cells captured.

Image of FIG. 9.
FIG. 9.

k value vs. effective antigen density of various channel segments, (a) with and (b) without bystander cells.

Image of FIG. 10.
FIG. 10.

Percent of cells captured as a function of bystander cell concentration.

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/content/aip/journal/bmf/5/3/10.1063/1.3623748
2011-08-22
2014-04-19
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752b84549af89a08dbdd7fdb8b9568b5 journal.articlezxybnytfddd
Scitation: Efficient capture of circulating tumor cells with a novel immunocytochemical microfluidic device
http://aip.metastore.ingenta.com/content/aip/journal/bmf/5/3/10.1063/1.3623748
10.1063/1.3623748
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