Microfluidic-Based Cell Sorting of Francisella tularensis Infected Macrophages Using Optical Forces
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- 作者：Thomas D. Perroud ; Julia N. Kaiser ; Jay C. Sy ; Todd W. Lane ; Catherine S. Branda ; Anup K. Singh ; Kamlesh D. Patel
- 刊名：Analytical Chemistry
- 出版年：2008
- 出版时间：August 15, 2008
- 年：2008
- 卷：80
- 期：16
- 页码：6365-6372
- 全文大小：341K
- 年卷期：v.80,no.16(August 15, 2008)
- ISSN：1520-6882
文摘
We have extended the principle of optical tweezers as a noninvasive technique to actively sort hydrodynamically focused cells based on their fluorescence signal in a microfluidic device. This micro fluorescence-activated cell sorter (µFACS) uses an infrared laser to laterally deflect cells into a collection channel. Green-labeled macrophages were sorted from a 40/60 ratio mixture at a throughput of 22 cells/s over 30 min achieving a 93% sorting purity and a 60% recovery yield. To rule out potential photoinduced cell damage during optical deflection, we investigated the response of mouse macrophage to brief exposures (<4 ms) of focused 1064-nm laser light (9.6 W at the sample). We found no significant difference in viability, cell proliferation, activation state, and functionality between infrared-exposed and unexposed cells. Activation state was measured by the phosphorylation of ERK and nuclear translocation of NF-κB, while functionality was assessed in a similar manner, but after a lipopolysaccharide challenge. To demonstrate the selective nature of optical sorting, we isolated a subpopulation of macrophages highly infected with the fluorescently labeled pathogen Francisella tularensis subsp. novicida. A total of 10 738 infected cells were sorted at a throughput of 11 cells/s with 93% purity and 39% recovery.