| This work focuses on the design and testing of a microchip that operates in a "flow cytometry-like" format. Microfluidic devices were used in combination with a fluorescence detection system. A multi-wavelength detection system was built and optimized conditions for data collection were obtained by adjusting LabView and Origin 7.0 software to suit the experimental conditions. We also developed an "in-house" procedure for isolation of mitochondria from Jurkat cells. Once the equipment was ready, we tested it by performing dose and time response studies, treating Jurkat cells and isolated mitochondria with valinomycin and oligomycin. Mitochondria membrane potential (DeltaPsi M) changes were monitored with JC-1 staining, which indicates changes in polarization of mitochondria membranes after drug treatment. The same type of studies was performed using the conventional technique, flow cytometry, and the results were compared with those obtained using the chip and multi-wavelength detection system. Our results demonstrate that microchip with a multi-wavelength detection system and flow cytometry gave similar results. |
