Biophysical Society 2020 Daily Schedule

Room 33A: Monday, February 17 9:30 AM – 11:00 AM Bruker Corporation

11:30 AM – 1:00 PM Leica Microsystems Leica SP8 FALCON: Applications of FLIM for Functional Imaging and STED Nanoscopy The rapidly growing field of functional imaging helps us understand the complex interactions of molecules, revealing the true nature of the underlying biology. In this context, fluorescence lifetime imaging (FLIM) is a powerful tool, providing valuable information beyond spectral imag- ing. FLIM is immune to concentration artifacts and highly sensitive to the molecular environment, providing a robust measure of a biological system’s health. However, previous FLIM solutions were slow and dif- ficult to implement, particularly for complex imaging workflows. To address this weakness, we present the Leica SP8 FALCON (Fast Lifetime Contrast), a fast, intuitive and totally integrated, all-Leica FLIM solution. The SP8 FALCON delivers video-rate FLIM with pixel-by-pixel quantifica- tion, due to a unique combination of fast electronics, sensitive spectral hybrid detectors (Leica HyDs), and a novel concept for measuring time. The system has ultra-short dead time and powerful built-in algorithms to manage data acquisition and analysis, while maintaining accuracy and excellent data quality. This talk explains the technical implementations enabling this new level of performance and provides some interesting application examples, including functional imaging (e.g. metabolic imaging or FRET imag- ing) and the use of lifetime information to achieve improved live-cell Nanoscopic Imaging ( τ- STED). τ -STED is a revolutionary modality for STED imaging, making use of the FALCON FLIM phasor approach, delivering cutting-edge resolution and image quality at low light dose, especially beneficial for live-cell nanoscopy applications. τ -STED takes the fluorescence lifetime information from all detected photons com- bined with phasor analysis in a novel way to increase the resolution and eliminate uncorrelated background in an automated manner. The τ- STED implementation on Leica SP8 STED 3x systems works for 2D and 3D STED in live and in fixed specimens, and for multicolor applications. The deep integration of SP8 FALCON into the Leica SP8 platformprovides easy access to complex FLIM experiments, enabling fast FLIM-FRET, 3D- and 4D-imaging modes, high-content screening, and auto-fluorescence component separation. Speaker Haridas Pudavar, Product Performance Manager-Confocal Systems, Leica Microsystems

From Single Molecules to Tissues – A New AFM Toolkit for Nanoscopic Investigation of Mechanics, Structures, and Dynamic Processes in Life Science The ability of atomic force microscopy (AFM) to obtain three-dimen- sional topography images of biological molecules and complexes with nanometer resolution and under near-physiological conditions remains unmatched by other imaging techniques. JPK BioAFM has developed a new NanoWizard® 4 XP AFM which not only enables the high-speed study of the time-resolved dynamics associated with cellular processes, it’s latest scanner technologies and compact design also allow full inte- gration of AFM into advanced commercially available light microscopy techniques. This seminar will focus on how the advances in Bruker’s latest BioAFM can be applied to study a wide-range of biological samples, from individual biomolecules to mammalian cells and tissues in real-time, in-situ experiments. We will present examples of how we are able to resolve the nanoscale structure of individual biomolecules at high-speed scan rates (150 Hz), follow the dynamic reorganization of the membrane-associated cytoskeleton of living cells at high-temporal and high-spatial resolution, and automatically map the topography of cell cultures across the entire area of the microscope stage. We will also discuss the full suite of BioAFM modes and accessories for studying the nanomechanical properties of cells and tissues, including direct correla- tion of multiparametric, quantitative AFM and super-resolution (STED) datasets. Speaker Andrea Slade, BioAFM Product Manager, Bruker Corporation

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