A fundamental need in almost all areas of biomedical research is the ability to separate single or small groups of cells from within a heterogeneous population. In order to obtain a living cell possessing a desired characteristic, individual cells or homogeneous groups of cells within the population must be analyzed followed by identification and isolation of the target cell(s). Most live-cell separation methods require that cells be dispersed into a single-cell suspension. Unfortunately, many crucial studies of cell biology cannot be performed in this manner. In the current application, an interdisciplinary team is collaborating to address the technical challenge of analyzing, sorting and collecting viable cells from a mixed population while they remain adherent to their growth surface. Easily implemented fabrication techniques and chemical surface modifications will be used to produce large arrays of releasable cell """"""""pallets"""""""" for the analysis, release and collection of single cells or colonies. Adherent cells will be cultured on the array, and the array will be analyzed using standard methods from image cytometry. Pallets containing single cells or small cell colonies will be individually released using a pulsed laser and collected for further analysis or expansion of the cells. Several applications for this system will be demonstrated. The early and rapid establishment of stably transfected cell lines based on the ability to identify and collect fluorescent microcolonies of cells expressing a red fluorescent protein fused with a key signal transduction enzyme will be performed. The value of selection based on morphology will be shown by cloning pure populations of cells transformed by an oncogenic retroviral protein. In addition, the approach enables living cells to be collected based on their dynamic characteristics, a selection criterion not feasible with current techniques. This capability will be used to select single cells based on siRNA perturbation of their calcium response or protein translocation. The expected benefits of this separation strategy are maintenance of cell viability, reduction in time and manipulation of the selected cells, and a broader set of cell attributes available for cell selection. The applications which involve basic cell biology, tumorigenesis, and signaling mechanisms in diabetes mellitus will demonstrate that the novel separation system should be widely applicable to the biomedical sciences.

National Institute of Health (NIH)
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Research Project (R01)
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Instrumentation and Systems Development Study Section (ISD)
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Korte, Brenda
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University of North Carolina Chapel Hill
Schools of Arts and Sciences
Chapel Hill
United States
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Pai, Jeng-Hao; Kluckman, Kimberly; Cowley, Dale O et al. (2013) Efficient division and sampling of cell colonies using microcup arrays. Analyst 138:220-8
Wang, Yuli; Shah, Pavak; Phillips, Colleen et al. (2012) Trapping cells on a stretchable microwell array for single-cell analysis. Anal Bioanal Chem 402:1065-72
Detwiler, David A; Dobes, Nicholas C; Sims, Christopher E et al. (2012) Polystyrene-coated micropallets for culture and separation of primary muscle cells. Anal Bioanal Chem 402:1083-91
Shadpour, Hamed; Zawistowski, Jon S; Herman, Annadele et al. (2011) Patterning pallet arrays for cell selection based on high-resolution measurements of fluorescent biosensors. Anal Chim Acta 696:101-7
Wang, Yuli; Dhopeshwarkar, Rahul; Najdi, Rani et al. (2010) Microdevice to capture colon crypts for in vitro studies. Lab Chip 10:1596-603
Wang, Yuli; Phillips, Colleen; Xu, Wei et al. (2010) Micromolded arrays for separation of adherent cells. Lab Chip 10:2917-24
Xu, Wei; Luikart, Alicia M; Sims, Christopher E et al. (2010) Contact printing of arrayed microstructures. Anal Bioanal Chem 397:3377-85
Gach, Philip C; Sims, Christopher E; Allbritton, Nancy L (2010) Transparent magnetic photoresists for bioanalytical applications. Biomaterials 31:8810-7
Shadpour, Hamed; Allbritton, Nancy L (2010) In situ roughening of polymeric microstructures. ACS Appl Mater Interfaces 2:1086-93
Xu, Wei; Sims, Christopher E; Allbritton, Nancy L (2010) Microcup arrays for the efficient isolation and cloning of cells. Anal Chem 82:3161-7

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