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Issue 3, 2013
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Laser-based directed release of array elements for efficient collection into targeted microwells

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Abstract

A cell separation strategy capable of the systematic isolation and collection of moderate to large numbers (25–400) of single cells into a targeted microwell is demonstrated. An array of microfabricated, releasable, transparent micron-scale pedestals termed pallets and an array of microwells in poly(dimethylsiloxane) (PDMS) were mated to enable selective release and retrieval of individual cells. Cells cultured on a pallet array mounted on a custom designed stage permitted the array to be positioned independently of the microwell locations. Individual pallets containing cells were detached in a targeted fashion using a pulsed Nd:YAG laser. The location of the laser focal point was optimized to transfer individual pallets to designated microwells. In a large-scale sort (n = 401), the accuracy, defined as placing a pallet in the intended well, was 94% and the collection efficiency was 100%. Multiple pallets were observed in only 4% of the targeted wells. In cell sorting experiments, the technique provided a yield and purity of target cells identified by their fluorescence signature of 91% and 93%, respectively. Cell viability based on single-cell cloning efficiency at 72 h post collection was 77%.

Graphical abstract: Laser-based directed release of array elements for efficient collection into targeted microwells

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Publication details

The article was received on 18 Sep 2012, accepted on 28 Nov 2012 and first published on 28 Nov 2012


Article type: Paper
DOI: 10.1039/C2AN36342A
Citation: Analyst, 2013,138, 831-838
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    Laser-based directed release of array elements for efficient collection into targeted microwells

    N. C. Dobes, R. Dhopeshwarkar, W. H. Henley, J. M. Ramsey, C. E. Sims and N. L. Allbritton, Analyst, 2013, 138, 831
    DOI: 10.1039/C2AN36342A

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