Biophysical Society 65th Annual Meeting Program Guide

1:30 PM – 2:00 PM Andor Technology

3:30 PM – 4:00 PM Oxford Instruments NanoAnalysis

SRRF-Stream+ - A Flexible and Effective Solution For Real-Time and Live Cell Super-Resolution Microscopy Much of the inner workings of the cell are hidden from view below the classical diffraction limit of light-based imaging microscopy. Super- resolution techniques such as STORM, PALM, STED and SIM have smashed past this barrier and have helped enable cell biology to be studied in considerably more detail. However, there are limitations to these techniques especially when considering live cell imaging. Super-resolution techniques may require costly microscopy equipment, high illumination intensities, long acqui- sition times or specialised fluorophores. SRRF (Super-resolution Radial Fluctuations) offers an alternative software-based approach that coun- ters many of these limitations (Gustafsson et al 2016). Specifically, it allows for super resolution at low illumination intensi- ties, using standard fluorophores on a conventional microscope. Understandably SRRF has become widely used. One development of SRRF is SRRF-Stream. This version is exclusive to Andor Technology and optimizes GPU processing to unlock real-time live super-resolution from a microscope. We now present an updated version of SRRF-Stream called “SRRF- Stream+” which allows for improvements in the image quality over the original version of SRRF-Stream. We also show that SRRF-Stream+ can be used on Andor Sona back-illuminated sCMOS cameras having previ- ously been available solely on Andor iXon EMCCD cameras. These new developments add to the previous benefits of SRRF-Stream, making it an even more flexible and useful part of the microscopists imaging toolbox. Speaker Alan Mullan, Product Application Specialist – Microscopy Cameras, Andor Technology

Multi-Colour Electron Microscopy: Using Energy Disper- sive X-ray Spectrometry to Image and Analyse Biological Samples The visualisation and analysis of life science samples has been a chal- lenge throughout the history of electron microscopy. Biological sample preparation and the absence or addition of contrasting agents often play a key role in the development of imaging methodology. But the signals generated in an electron microscope are mostly underutilised by biologists. While energy dispersive x-ray spectrometry (EDS) has been used in materials science for many decades, sample stability and detector sensitivity have prevented a broader adoption in life sciences until recently [1]. Multi-colour electron microscopy (MCEM) combines elemental information about samples produced using EDS with ultra- structural electron data, providing a powerful and informative imaging technique [2]. MCEM addresses key research topics for the biological electron micros- copist. What is it, where is it and how much? Using examples from bio- medical research, animal cells and tissues, and plant cell biology, this talk will demonstrate how the addition of elemental maps to electron images contributes key information that could be used for a variety of biological imaging applications, such as region of interest profiling or automated segmentation of volumetric data). EDS is not only a power- ful imaging tool, providing accurate identification of stains, labels, and ultrastructural features, but it can also be used to conduct analysis on the relative quantities of a wide range of elements, providing composi- tional data on native elements and exogenous features. Speaker Louise Hughes, Product Manager Life Science, Oxford Instruments [1] Pirozzi, N.M., Hoogenboom, J.P. and Giepmans, B.N., 2018. ColorEM: analytical electron microscopy for element-guided identifica- tion and imaging of the building blocks of life. Histochemistry and cell biology, 150(5), pp.509-520. [2] Scotuzzi, M., Kuipers, J., Wensveen, D.I., De Boer, P., Hoogenboom, J.P. and Giepmans, B.N., 2017. Multi-color electron microscopy by ele- ment-guided identification of cells, organelles and molecules. Scientific reports, 7(1), pp.1-8.

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