Planar silicon chips with 1-2-microm etched holes (average resistance: 2.04 +/- 0.02 MOmega in physiological buffer, n = 274) have been developed for patch-clamp recordings of whole-cell currents from cells in suspension. An automated 16-channel parallel screening system, QPatch 16, has been developed using this technology. A single-channel prototype of the QPatch system was used for validation of the patch-clamp chip technology. We present here data on the quality of patch-clamp recordings and from actual drug screening studies of human potassium channels expressed in cultured cell lines. Using Chinese hamster ovary (CHO) and human embryonic kidney cells (HEK), gigaseals of 4.1 +/- 0.4 GOmega (n = 146) and high-quality whole-cell current recordings were obtained from hERG and KCNQ4 potassium channels. Success rates for gigaseal recordings varied from 40 to 95%, and 67% of the whole-cell configurations lasted for >20 min. Cells were maintained in suspension up to 4 h in a cell storage facility that is integrated in the QPatch 16. No decline in patchability was observed during this time course. A series of screens was conducted with known inhibitors of the hERG and KCNQ4 potassium channels. Dose-response relationship characterizations of verapamil and rBeKm-1 blockage of hERG currents provided IC(50) values similar to values reported in the literature.
We demonstrate efficient four-wave mixing among different spatial modes in a 1-km long two-mode fiber at telecommunication wavelengths. Two pumps excite the LP 01 and LP 11 modes, respectively, while the probe signal excites the LP 01 mode, and the phase conjugation (PC) and Bragg scattering (BS) idlers are generated in the LP 11 mode. For these processes we experimentally characterize their phase matching efficiency and bandwidth and find that they depend critically on the wavelength separation of the two pumps, in good agreement with the numerical study we carried out. We also confirm experimentally that BS has a larger bandwidth than PC for the optimum choice of the pump wavelength separation.
Diatoms are ubiquitous organisms in aquatic environments and are estimated to be responsible for 20-25 % of the total global primary production. A unique feature of diatoms is the silica wall, called the frustule. The frustule is characterized by species-specific intricate nanopatterning in the same size range as wavelengths of visible and ultraviolet (UV) light. This has prompted research into the possible role of the frustule in mediating light for the diatoms' photosynthesis as well as into possible photonic applications of the diatom frustule. One of the possible biological roles, as well as area of potential application, is UV protection. In this review, we explore the possible adaptive value of the silica frustule with focus on research on the effect of UV radiation on diatoms. We also explore the possible effect of the frustules on UV radiation, from a theoretical, biological, and applied perspective, including recent experimental data on UV transmission of diatom frustules.
Effective screening of large compound libraries in ion channel drug discovery requires the development of new electrophysiological techniques with substantially increased throughputs compared to the conventional patch clamp technique. Sophion Bioscience is aiming to meet this challenge by developing two lines of automated patch clamp products, a traditional pipette-based system called Apatchi-1, and a silicon chip-based system QPatch. The degree of automation spans from semi-automation (Apatchi-1) where a trained technician interacts with the system in a limited way, to a complete automation (QPatch 96) where the system works continuously and unattended until screening of a full compound library is completed. The performance of the systems range from medium to high throughputs.
Automated patch clamp (APC) devices have become commonplace in many industrial and academic labs. Their ease-of-use and flexibility have ensured that users can perform routine screening experiments and complex kinetic experiments on the same device without the need for months of training and experience. APC devices are being developed to increase throughput and flexibility. Areas covered: Experimental options such as temperature control, internal solution exchange and current clamp have been available on some APC devices for some time, and are being introduced on other devices. A comprehensive review of the literature pertaining to these features for the Patchliner, QPatch and Qube and data for these features for the SyncroPatch 384/768PE, is given. In addition, novel features such as dynamic clamp on the Patchliner and light stimulation of action potentials using channelrhodosin-2 is discussed. Expert opinion: APC devices will continue to play an important role in drug discovery. The instruments will be continually developed to meet the needs of HTS laboratories and for basic research. The use of stem cells and recordings in current clamp mode will increase, as will the development of complex add-ons such as dynamic clamp and optical stimulation on high throughput devices.
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