Cytosolic Calcium Measurement Utilizing a Single-Cell Biochip to Study the Effect of Curcumin and Resveratrol on a Single Glioma Cell.

A microfluidic method has been developed for real-time measurement of the effects of curcumin on the intracellular calcium concentration in a single glioma cell (U87-MG). This method is based on quantitative fluorescence measurement of intracellular calcium in a cell selected in a single-cell biochip. This biochip consists of three reservoirs, three channels, and a V-shaped cell retention structure. Because of the adherent nature of glioma cells, a single cell can adhere within the aforementioned V-shaped structure. The single-cell calcium measurement will minimize cell damage caused by conventional cell calcium assay methods. Previous studies have shown that curcumin increased cytosolic calcium in glioma cells using the fluorescent dye: Fluo-4. So in this study, the effects of 5 µM and 10 µM solutions of curcumin on the increases of cytosolic calcium in a single glioma cell have been measured. Moreover, the effects of 100 µM and 200 µM of resveratrol are measured. At the final stage of the experiments, ionomycin was used to increase the intracellular calcium to the highest possible level due to dye saturation. It has been demonstrated that microfluidic cell calcium measurement is a real-time cytosolic assay that requires small quantities of reagent, which will have potential uses for drug discovery.

The microfluidic capture of single breast cancer cells for multi-drug resistance assays.

Utilizing the microfluidic single-cell technique enables us to study the inhibition of multidrug resistance due to drug efflux on a single triple-negative breast cancer cell. This method examines drug efflux inhibition on a single cell in a microfluidic chip using a conventional optical detection system constructed from an inverted microscope and a microphotometer. More importantly, the integration of single-cell selection, dye and drug loading, and fluorescence measurement for intracellular drug accumulation is all conducted on a single microfluidic chip. By using a microfluidic chip and the adherent nature of the cancer cell lines, a single breast cancer cell could be selected and retained near the cell retention structure in the chip. This enabled us to detect dye accumulation in the MDR breast cells in the presence of cyclosporine A (CsA). CsA and rhodamine 123 (Rh123) were used as the P-glycoprotein (P-gp) inhibitor and fluorescent dye, respectively. Furthermore, Paclitaxel, a commonly known chemotherapeutic used in breast cancer patients, was administered in the presence of both reagents. During the entirety of the experiment fluorescence measurement was used to monitor the fluctuating levels of intracellular Rh123 levels, and an optical imaging system was used to monitor the shape and size of the cell. The results showed that the Rh123 fluorescence signal in a single-cell increased dramatically over its same-cell control due to the competitive inhibition of paclitaxel and the non-competitive inhibition subjected by CsA.