Abstract
Conventional cell trapping methods using microwells with small dimensions (10–20 μm) are useful for examining the instantaneous cell response to reagents; however, such wells have insufficient space for longer duration screening tests that require observation of cell attachment and division. Here we describe a flow method that enables single cell trapping in microwells with dimensions of 50 μm, a size sufficient to allow attachment and division of captured cells. Among various geometries tested, triangular microwells were found to be most efficient for single cell trapping while providing ample space for cells to grow and spread. An important trapping mechanism is the formation of fluid streamlines inside, rather than over, the microwells. A strong flow recirculation occurs in the triangular microwell so that it efficiently catches cells. Once a cell is captured, the cell presence in the microwell changes the flow pattern, thereby preventing trapping of other cells. About 62% of microwells were filled with single cells after a 20 min loading procedure. Human prostate cancer cells (PC3) were used for validation of our system.
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Acknowledgments
The authors would like to thank Chentian Zhang for assisting experiments, and Dr. Rachel Schmedlen for supporting a student team consisting of five authors who contributed equally to this study: M. Morgan, A. N. Sachs, J. Samorezov, R. Teller, and Y. Shen. This study was supported by the Wilson Foundation, Coulter Foundation, and the UMCCC Prostate SPORE P50 CA69568 pilot grant. Dr. J. Y. Park was supported by the Korea Research Foundation Grant, Republic of Korea (KRF-2008-357-D00030). Dr. K. J. Pienta is supported by NIH Grant PO1 CA093900, an American Cancer Society Clinical Research Professorship, NIH SPORE in prostate cancer Grant P50 CA69568, and the Cancer Center support Grant P30 CA46592. This work was supported in part by a generous grant from Mr. and Mrs. Turner.
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Park, J.Y., Morgan, M., Sachs, A.N. et al. Single cell trapping in larger microwells capable of supporting cell spreading and proliferation. Microfluid Nanofluid 8, 263–268 (2010). https://doi.org/10.1007/s10404-009-0503-9
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DOI: https://doi.org/10.1007/s10404-009-0503-9