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Nat Biotechnol ; 23(1): 83-7, 2005 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-15608628

RESUMO

Microfluidic-based devices have allowed miniaturization and increased parallelism of many common functions in biological assays; however, development of a practical technology for microfluidic-based fluorescence-activated cell sorting has proved challenging. Although a variety of different physical on-chip switch mechanisms have been proposed, none has satisfied simultaneously the requirements of high throughput, purity, and recovery of live, unstressed mammalian cells. Here we show that optical forces can be used for the rapid (2-4 ms), active control of cell routing on a microfluidic chip. Optical switch controls reduce the complexity of the chip and simplify connectivity. Using all-optical switching, we have implemented a fluorescence-activated microfluidic cell sorter and evaluated its performance on live, stably transfected HeLa cells expressing a fused histone-green fluorescent protein. Recovered populations were verified to be both viable and unstressed by evaluation of the transcriptional expression of two genes, HSPA6 and FOS, known indicators of cellular stress.


Assuntos
Separação Celular/métodos , Citometria de Fluxo/métodos , Microfluídica/métodos , Análise de Sequência com Séries de Oligonucleotídeos , Tamanho Celular , Proteínas de Fluorescência Verde/química , Proteínas de Fluorescência Verde/metabolismo , Células HeLa , Histonas/química , Humanos , Lasers , Polipropilenos/química , RNA Mensageiro/metabolismo , Semicondutores , Espectrometria de Fluorescência/métodos , Temperatura , Transcrição Gênica , Transfecção
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