Microfluidic-Based Cell Sorting of Francisella tularensis Infected Macrophages Using Optical Forces

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 co...

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Published inAnalytical chemistry (Washington) Vol. 80; no. 16; pp. 6365 - 6372
Main Authors Perroud, Thomas D, Kaiser, Julia N, Sy, Jay C, Lane, Todd W, Branda, Catherine S, Singh, Anup K, Patel, Kamlesh D
Format Journal Article
LanguageEnglish
Published Washington, DC American Chemical Society 15.08.2008
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Summary: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.
Bibliography:istex:A4D63D27483FF9E8C25ABE74DC9BE832A76D79EF
Video of single macrophages optically deflected by an IR laser in a microfluidic chip and Figures S-2 and S-3. This material is available free of charge via the Internet at http://pubs.acs.org.
ark:/67375/TPS-QZHPW6SP-W
ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
SAND2008-2737J
USDOE National Nuclear Security Administration (NNSA)
DE-AC04-94AL85000
ISSN:0003-2700
1520-6882
DOI:10.1021/ac8007779