Chang Yu Chen

Trapping of Bioparticles via Microvortices in a Microfluidic Device for Bioassay Applications

This paper presents hydrodynamic trapping of bioparticles in a microfluidic device. An in-plane oscillatory microplate, driven via Lorentz law, generates two counter-rotating microvortices, trapping the bioparticles within the confines of the microvortices. The force required to trap bioparticles is quantified by tuning the background flow and the microplates excitation voltage. Trapping and releasing of 10-microm polystyrene beads, human embryonic kidney (HEK) cells, red blood cells (RBCs), and IgG antibodies were demonstrated. Results show the microvortices rotates at 0-6 Hz corresponding to 2-9 Vpp (peak-to-peak) excitation. At a particular rate of rotation (2-7 Vpp tested), a bioparticle is trapped until the background flow exceeds a limit. This flow limit increases with the rate of rotation, which defines the trap/release force boundary over the range of operation. This boundary is 12 +/- 2.0 pN for cell-size bioparticles and 160 +/- 50 fN for antibodies. Trapping of RBCs demonstrated microvortices ability for nonspherical cells. Cell viability was studied via HEK cells that were trapped for 30 min and shown to be viable. This hydrodynamically controlled approach to trap a wide range of bioparticles should be useful as a microfluidic device for cellular and subcellular bioassay applications.

Patch clamping on plane glass—fabrication of hourglass aperture and high-yield ion channel recording

Planar patch-clamp has revolutionized ion-channel measurement by eliminating laborious manipulation from the traditional micropipette approach and enabling high throughput. However, low yield in gigaseal formation and/or relatively high cost due to microfabricated processes are two main drawbacks. This paper presents patch clamping on glass substrate-an economical solution without sacrificing gigaseal yield rate. Two-stage CO(2) laser drilling methodology was used to generate an hourglass, funnel-like aperture of a specified diameter with smooth and debris-free surfaces on 150 microm borosilicate cover glass. For 1-3 microm apertures as patch-clamp chips, seal resistance was tested on human embryonic kidney, Chinese hamster ovary, and Jurkat T lymphoma cells with a gigaseal success rate of 62.5%, 43.6% and 66.7% respectively. Results also demonstrated both whole-cell and single channel recording on endogenously expressed ion channels to confirm the capability of different patch configurations.

Ion channel electrophysiology via integrated planar patch-clamp chip with on-demand drug exchange

Planar patch clamp has revolutionized characterization of ion channel behavior in drug discovery primarily via advancement in high throughput. Lab use of planar technology, however, addresses different requirements and suffers from inflexibility to enable wide range of interrogation via a single cell. This work presents integration of planar patch clamp with microfluidics, achieving multiple solution exchanges for tailor‐specific measurement and allowing rapid replacement of the cell‐contacting aperture. Studies via endogenously expressed ion channels in HEK 293T cells were commenced to characterize the device. Results reveal the microfluidic concentration generator produces distinct solution/drug combination/concentrations on‐demand. Volume‐regulated chloride channel and voltage‐gated potassium channels in HEK 293T cells immersed in generated solutions under various osmolarities or drug concentrations show unique channel signature under specific condition. Excitation and blockage of ion channels in a single cell was demonstrated via serial solution exchange. Robustness of the reversible bonding and ease of glass substrate replacement were proven via repeated usage of the integrated device. The present approach reveals the capability and flexibility of integrated microfluidic planar patch‐clamp system for ion channel assays. Biotechnol. Bioeng. 2011; 108:1395–1403.