Biophysical Society 66th Annual Meeting Program Guide

Exhibitor Presentations Esplanade, Rooms 157 and 158, Moscone Center

Esplanade, Room 157: Sunday, February 20 11:30 AM – 1:00 PM Carl Zeiss Microscopy LLC The ZEISS LSM 9 Series – Redefining Your Confocal Experi- ence The confocal experience continues to evolve with the new features available on the ZEISS LSM 9 Series. As your model systems continue to get larger in order to address the relevant physiological questions, we continue to push the limits of speed, resolution, and sensitivity, as well as spectral flexibility to meet these demands. This includes our revolutionary Airyscan 2 technology, the most sensitive NIR detection available, and now advanced, but easy to use processing tools such as LSM Plus, to ensure that you obtain the best image quality at any bud- get. Furthermore, we now talk about entire workflows by incorporating AI based tools both for acquisition and analysis. From images to results, our powerful ZEN software has unique, efficient, and easy to use strat- egies to ensure you have the best images and data possible as well as sophisticated, but user-friendly data and image analysis, including many options for machine learning and automation. Join us in learning about the latest new software and hardware features available on the ZEISS LSM 9 Series confocal microscope systems. Speaker Samantha Fore, Product Marketing Manager Laser Scanning Microscopy Life Sciences, Carl Zeiss Microscopy LLC 1:30 PM – 3:00 PM Mad City Labs Inc Novel Single Molecule Methods for Tracking, Transport, and Protein Complex Analyses Capturing the Early Stages of the Virus-Cell Interaction With Active- Feedback Single-Virus Tracking Kevin Welsher

Measuring Activity of Single Transporters in Single Vesicles Using TIRF Microscopy Joseph Mindell Secondary active transporters are essential contributors to ion and substrate fluxes across biological membranes. One limitation to our current knowledge is that transporter activity measurements generally reflect the average behavior of a multitude of proteins, while the indi- vidual behaviors of single transporters are unresolved. Here we report development of a novel, single-liposome assay using single-molecule techniques to measure the activity of individual transporters using ClC-ec1, a member of the CLC family of Cl-/H+ exchangers as a model system. Using this method, we observe transport catalyzed by single CLC-ec1 dimers and investigate the nature of stoichiometric coupling in this antiporter. Rapid Extraction and Kinetic Analysis of Protein Complexes From Single Cells Sena Sarıkaya & Dan Dickinson Understanding assembly of molecular machines in developing cells requires a quantitative, biochemical, and in vivo approach. We devel- oped an assay to interrogate the abundance, dynamics, and stability of native protein complexes extracted from single cells. We optically lysed Caenorhabditis elegans zygotes, captured in vivo single protein com- plexes on a coverslip using antibodies, and monitored the dynamics of these complexes over time using multi-color MicroMirror Total Internal Reflection Fluorescence microscopy. We developed open-source soft- ware to process thousands of kinetic measurements per cell. This assay provides an unprecedented level of resolution as compared to tradi- tional in vitro biochemistry and can be applied to any protein pair that can be labeled and detected. Speakers Joseph Mindell, National Institute of Neurological Disorders and Stroke, NIH Sena Sarıkaya, Department of Molecular Biosciences, The University of Texas at Austin Kevin Welsher, Department of Chemistry, Duke University

target fluorophores in the focal volume of an optical microscope using real-time feedback to move the sample and compensate for molecular diffusion. 3D-SMART has been successfully applied to single proteins and nucleic acids

Here we introduce an active- feedback 3D microscopy tech- nique capture the dynamics of rapidly diffusing single molecules in solution (3D Single-Molecule Active Real-time Tracking or 3D-SMART). This method “locks"

at diffusive speeds up to 10 μm 2/s. We will further describe how this microscope, when combined with rapid volumetric imaging, can cap- ture the early events in the interactions between single viral particles and live cells in three dimensions with millisecond or better temporal resolution.

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