Abstract
Bilayer Lipid Membranes (BLM) are often used to perform single-channel recordings of different ion-channels. However, the success ratio and the BLM stability is strongly dependent on the specific electrophysiology setup, which is usually bulky and expensive. Moreover, the post-processing acquisition analysis is a time-consuming activity. These characteristics make BLM experiments difficult for educational purposes.Here we present a compact and complete BLM educational kit for accurate electrophysiological recordings allowing fast screening of ion-channels functional activity in lipid environments. A large number of standard experiments can be performed using the kit, which is particularly suitable for educational lab courses. Stable membranes can be easily formed to study ion-channels or toxins.The real-time statistic feature implemented in the amplifier control software enables quick and straightforward analysis of data about ion-channel properties, such as I/V curve for conductance analysis, FFT, dwell times, histograms and inter-events time statistics.The kit is contained into a 16x10x5cm Faraday cage and includes the recording chamber and the miniaturized amplifier. The chamber is based on two 1ml - 30x15x15mm Delrin cuvettes with individual electrodes. A laser drilled Teflon diaphragm, with holes varying between 50µm to 110µm, separates the two cuvettes.To prove the capability of the presented setup and of the real-time statistics, suspended lipid bilayer were easily formed by painting method and recordings of different proteins, such as Kcv, Gramicidin and alpha-hemolysin in presence of PEGs of different lengths, were analyzed. Thanks to the real-time statistics, scientists can perform a quick analysis on the ion-channel properties while experiments are running.The combination of the presented kit and the control software with real-time statistics feature allows quick and easy functional characterizations of many kinds of ion-channels for educational or research purposes.
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