Laser-based directed release of array elements for efficient collection into targeted microwells
| Autor: | Nancy L. Allbritton, Nicholas C. Dobes, W. Hampton Henley, Rahul Dhopeshwarkar, Christopher E. Sims, J. Michael Ramsey |
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| Rok vydání: | 2013 |
| Předmět: |
Materials science
Cell Survival Green Fluorescent Proteins Nanotechnology Tissue Array Analysis Cell Separation Transfection Biochemistry Article Analytical Chemistry law.invention law Cell Line Tumor Electrochemistry Cell separation Humans Environmental Chemistry Dimethylpolysiloxanes Pallet Spectroscopy Cell survival Lasers Laser HeLa Cells Biomedical engineering |
| Zdroj: | The Analyst. 138:831-838 |
| ISSN: | 1364-5528 0003-2654 |
| DOI: | 10.1039/c2an36342a |
| Popis: | 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%. |
| Databáze: | OpenAIRE |
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