Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology; Center for Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital
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Chung, B. G., Manbachi, A., Khademhosseini, A. A Microfluidic Device with Groove Patterns for Studying Cellular Behavior. J. Vis. Exp. (7), e270, doi:10.3791/270 (2007).
We describe a microfluidic device with microgrooved patterns for studying cellular behavior. This microfluidic platform consists of a top fluidic channel and a bottom microgrooved substrate. To fabricate the microgrooved channels, a top poly(dimethylsiloxane) (PDMS) mold containing the impression of the microfluidic channels was aligned and bonded to a microgrooved substrate. Using this device, mouse fibroblast cells were immobilized and patterned within microgrooved substrates (25, 50, 75, and 100 μm wide). To study apoptosis in a microfluidic device, media containing hydrogen peroxide, Annexin V, and propidium iodide was perfused into the fluidic channel for 2 hours. We found that cells exposed to the oxidative stress became apoptotic. These apoptotic cells were confirmed by Annexin V that bound to phosphatidylserine at the outer leaflet of the plasma membrane during the apoptosis process. Using this microfluidic device with microgrooved patterns, the apoptosis process was observed in real-time and analyzed by using an inverted microscope containing an incubation chamber (37°C, 5% CO2). Therefore, this microfluidic device incorporated with microgrooved substrates could be useful for studying the cellular behavior and performing high-throughput drug screening.
A. Microfabrication of the microfluidic device
B. Assembling the device
C. Cell seeding and experimental setup
Cells were immobilized and patterned within microgrooved substrates in a microfluidic device. The apoptosis process of cells exposed to hydrogen peroxide was observed in real-time and analyzed by using Annexin V and propidium iodide. Thus, this microfluidic device containing microgroove channels could be useful for high-throughput drug screening.
|PDMS||Reagent||K.R. Anderson Co.||2065622||Poly(dimethylsiloxane), Dow Corning Sylgard 184 (8.6 lb)|
|DMEM||medium||Invitrogen||11965||Dulbecco’s Modified Eagle’s Media|
|FBS||serum||Invitrogen||10082-147||Fetal Bovine Serum|
|Apoptosis assay||Invitrogen||V13242||Annexin A, propidium iodide|
|Negative photoresist||MicroChem Corp.||SU-8 2015|
|Si wafer||Tool||4 inch silicone wafer|
|Reactive oxygen plasma||Reagent||Harrick Scientific Products, Inc.||treat wafer 5 min at 30W|
|inverted microscope||Tool||Nikon Instruments||TE 2000|
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