Research Papers: Sensing

Microfluidic approach for direct and uniform laser irradiation to study biochemical state changes on Jurkat-T cells

[+] Author Affiliations
Sween J. Butler

University of North Texas, Department of Physics, 1155 Union Circle, #311427, Denton, Texas 76203

Dong Weon Lee

Chonnam National University, School of Mechanical Systems Engineering, Gwang-Ju 500-757, Republic of Korea

Curtis W. Burney

United States Air Force Academy, Department of Biology, 2355 Faculty Drive, Suite 2P483, Colorado 80840-6226

Jeffrey C. Wigle

Optical Radiation Bioeffects Branch, 4141 Petroleum Road, JBSA Fort Sam Houston, Texas 78234-2644

Tae Youl Choi

University of North Texas, Department of Mechanical and Energy Engineering, 3940 North Elm Street, Suite F-101J, Denton, Texas 76207

J. Biomed. Opt. 18(11), 117004 (Nov 06, 2013). doi:10.1117/1.JBO.18.11.117004
History: Received July 16, 2013; Revised October 10, 2013; Accepted October 14, 2013
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Abstract.  We investigated the potential for using polydimethylsiloxane microfluidic devices in a biological assay to explore the cellular stress response (CSR) associated with hyperthermia induced by exposure to laser radiation. In vitro studies of laser-tissue interaction traditionally involved exposing a monolayer of cells. Given the heating-cooling dynamics of the cells and nutrient medium, this technique produces a characteristic “bulls-eye” temperature history that plagues downstream molecular analyses due to the nonuniform thermal experience of exposed cells. To circumvent this issue, we devised an approach to deliver single cells to the laser beam using a microfluidic channel, allowing homogeneous irradiation and collection of sufficient like-treated cells to measure changes in CSR after laser heating. To test this approach, we irradiated Jurkat-T cells with a 2-μm-wavelength laser in one branch of a 100-μm-wide bifurcated channel while unexposed control cells were simultaneously passing through the other, identical channel. Cell viability was measured using vital dyes, and expression of HSPA1A was measured using reverse transcription polymerase chain reaction. The laser damage threshold was 25±2J/cm2, and we found a twofold increase in expression at that exposure. This approach may be employed to examine transcriptome-wide/proteome changes and further comparative work across stressors and cell types.

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© 2013 Society of Photo-Optical Instrumentation Engineers

Citation

Sween J. Butler ; Dong Weon Lee ; Curtis W. Burney ; Jeffrey C. Wigle and Tae Youl Choi
"Microfluidic approach for direct and uniform laser irradiation to study biochemical state changes on Jurkat-T cells", J. Biomed. Opt. 18(11), 117004 (Nov 06, 2013). ; http://dx.doi.org/10.1117/1.JBO.18.11.117004


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