Biophysical Society 65th Annual Meeting Program Guide

Thursday, February 25 11:30 AM – 12:00 PM Nanion Technologies Automated Electrophysiology For Any Kinetics: Ion Chan- nels & Transporters Ion channels and transporters are important physiological and phar- macological targets. Electrophysiology remains the gold standard for studying these important targets and automation of the technique ensures higher throughput is achieved whilst maintaining high data quality. In this virtual symposium, Nanion Technologies provides two case studies where automated patch clamp (APC) or solid-supported membrane (SSM)-based electrophysiology devices were used in dif- ferent applications. After a short greeting by Dr. Niels Fertig (CEO), Nanion Technologies will welcome two exceptional speakers, Dr Nina Braun (University of Copenhagen) and Dr. Matthias Quick (Columbia University). Dr. Nina Braun presents recent work, with focus on establishing a high-throughput protocol to conduct functional and pharmacological investigations of non-canonical amino acids (ncAA)-containing hASIC1a (human acid-sensing ion channel 1a) variants in transiently transfected mammalian cells. Incorporation of ncAAs can endow proteins with novel functionalities, such as crosslinking or fluorescence. Function of these variants in ion channels can be studied with great precision using standard electrophysiology, but this approach is typically labor intensive and low throughput. During the study, three different pho- tocrosslinking ncAAs were introduced into 103 positions and the func- tion of the resulting 309 variants was assessed with SyncroPatch 384i automated patch-clamp platform, demonstrating that the approach is efficient and versatile, as it is amenable to assessing even complex pharmacological modulation by peptides. The data show that the acidic pocket is a major determinant for current decay and live-cell cross- linking provides insight into the hASIC1a-psalmotoxin-1 interaction. Overall, this protocol aims to enable future APC-based studies of ncAA- containing ion channels in mammalian cells. Next, Dr. Matthias Quick is focusing on the study of ion-dependent transporters with special emphasis on Na+ or H+-coupled symport- ers. Whereas flux studies with radiolabeled solutes use the target protein reconstituted in proteoliposomes provided a wealth of infor- mation, the determination of the thermodynamically-coupled solute transport-associated flux of H+ or Na+ has been challenging. By using the SURFE2R N1 SSM platform, his team was able to quickly collect data of solute transport-associated flux of co-transported ions across the membrane of proteoliposomes containing different target proteins. Additionally, with SURFE2R technology it is possible to collect data for a full kinetic characterization of a target protein such as its dependence on substrate and ion concentrations, pH, and potential essential addi- tives, as well as its substrate recognition profile. The SURFE2R system also enables the use of a wide range of substrates that are readily com- mercially available, avoiding the use of radiolabeled compounds. Speakers Nina Braun, Post-Doctoral Fellow, Department of Drug Design and Pharmacology, University of Copenhagen Matthias Quick, Associate Professor of Neurobiology (Psychiatry), Columbia University Medical Center (CUMC)

4:00 PM – 4:30 PM Carl Zeiss Microscopy LLC Discovering the Subcellular Dynamics of Life with ZEISS Lat- tice Lightsheet 7 In order to best understand the world around us it is necessary to observe microscopic specimins in as natural a state as possible. This requires a transition from imaging fixed to live specimins and expand- ing from 2D to 3D model organisms. The drive towards live-cell imaging over long timeframes and at high volume speeds brings new challenges. There is evidence that traditional imaging techniques can influence the behaviour of specimins due to phototoxicity, thus affecting the integrity of the results. The most influential technological breakthroughs which address these challenges have been modifications to the shape of the excitation light. Classical laser-based imaging approaches utilize a gaussian excitation beam which is focussed to a spot or a sheet and scanned as required to excite the sample. As an alternative approach, bessel beams have been combined to introduce a structured pattern to the beam profile. The resulting ‘lattice’ of light has many benefits for live imaging. The most notable are a reduction of light exposure due to significant improve- ment in signal to noise while maintaining high resolution and optical sectioning. With lattice-lightsheet microscopy it is possible to capture dynamics at previously unreachable combinations of acquisition speed and resolution over hours and even days. This talk will describe how the ZEISS Lattice Lighsheet 7 makes long- term volumetric imaging of living cells with subcellular resolution possible without having to change your standard sample preparation protocols to accomdate the instrument. With automatic alignment and easy acquisition workflows, lattice light-sheet imaging is now as acces- sible as using a standard inverted microscope. Join us for this webinar to learn how ZEISS Lattice Lightsheet 7 allows you to discover the subcellular dynamics of life. Speaker Renée Dalrymple, Product Marketing Manager – Life Sciences Lattice Line, Carl Zeiss Microscopy LLC

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