Biophysical Society 65th Annual Meeting Program Guide

Exhibitor Presentations

Tuesday February 23 11:30 AM – 12:00 PM Mad City Labs Inc From iSCAT to SCATTIRSTORM: Adventures in Single-Mole- cule Microscopy I will describe the design and application of two multi-modal micro- scopes built around the Mad City Labs RM21® single molecule micro- scope. The first system combines Interferometric Scattering (iSCAT) microscopy, developed by Philipp Kukura’s lab, together with Total Internal Reflection Dark-Field (TIRDM). By labeling kinesin-1 motors on one head with a 30-nm gold nanoparticle, we were able to track motor stepping at 1 kHz temporal resolution and 1-2 nm spatial preci- sion. This allowed us to clarify previously uncovered features of kinesin stepping. In collaboration with Luke Rice, we extended this system to understanding microtubule dynamics by labeling tubulin dimers with 20-nm gold nanoparticles and observing these tubulin reversibly bind- ing at the plus-ends of growing microtubules. These tools have the potential to be applied broadly to single-molecule studies to increase the temporal resolution beyond what is traditionally accessible by fluorescence, while still maintaining the spatial resolution provided by point-spread function fitting. In the second part of the talk, I will describe our progress on a multi-modal microscope that we are using for a DOE-funded project to study the mechanism of cellulose degra- dation by cellulases for bioenergy applications. In particular, we have optimized the ability to combine Interference Reflection Microscopy (IRM) to image cellulose micro- and nanofibers with TIRF to simultane- ously image fluorescently-labeled cellulase enzymes degrading the cellulose. Speaker William Hancock, Department of Biomedical Engineering, Pennsylvania State University

1:30 PM – 2:00 PM Bruker

Introduction to Super-Resolution Multiplexed Imaging Ap- plications Using the Vutara VXL Platform For Single-Mole- cule and Widefield Imaging This talk will introduce the next generation of the Vutara imaging plat- form, the VXL. While the VXL has been designed and optimized for sin- gle molecule localization microscopy, most current methods for super resolution microscopy are limited due to the limited number of dyes compatible with single molecule based super resolution techniques. This talk presents methods for imaging a series of consecutive targets within a sample using single molecule localization microscopy inte- grated with a software-controlled automated microfluidics system for probe multiplexing. Probe multiplexing allows for the imaging of more than four different targets within a cell. During this talk we will show examples using oligoSTORM and DNA-PAINT methods. OligoSTORM allows for the direct tracing of chromosomes within cells. We will show the three-dimensional trajectory of a multiplexed oligoPAINT labeled chromosome in individual human fibroblast cells using the Vutara platform. We will also show DNA-PAINT based single molecule localiza- tion data for antibody labeled targets in cell culture. We will also show how the Vutara can be used for ORCA, a widefield imaging technique that has been developed for high throughput sequential labelling of chromosome targets for generating 3D image data and single cell chromosome conformation maps (similar to Hi-C type data). The VXL with integrated fluidics and SRX software provides a powerful suite of tools for simultaneous imaging, localization, visualization and statistical analysis of multiplexed data. Speaker Robert Hobson, Applications Scientist, Bruker 3:30 PM – 4:00 PM Nikon Instruments Inc Using the Nikon Ti2 Inverted Platform as an Optical Bench Flexibility and modularity are core principles underlying Nikon’s imag- ing systems and components. In this presentation, we will discuss innovative solutions from Nikon that are designed to maximize flex- ibility and enable easy customization of both home-built microscopes and fully turn-key commercial systems. Learn how the Eclipse Ti2 with its expandable structure, multiple input and output ports, and easy access to the back-aperture plane can be used as an optical bench. Furthermore, Nikon’s wide range of modular illumination devices from TIRF to photo-stimulation can be custom-combined for your unique application. For those requiring full access to the optical path, Nikon also offers the Ti2-Hardware Development Kit for introducing com- pletely custom components. We will also discuss Nikon’s latest objec- tives utilizing new immersion media and featuring industry-leading numerical aperture and working distance. Speaker Ian Ross, Senior Biosystems Applications Manager, Nikon Instruments Inc

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