This proposal seeks funding for a Gatan Inc., OneView scintillator- coupled sCMOS transmission electron microscope (TEM) camera, a OneView 16bit computer with a Windows 10 operating system, and Gatan Microscopy Suite V3 software. This new sCMOS TEM camera will replace and upgrade the Stanford University Cell Sciences Imaging Facility?s Electron Microscopy Core?s (EMC) aging and obsolete CCD TEM camera. Every molecular, cell and tissue biomedical research program has needs for microscopic visualization of research specimens. For many biomedical research projects this visualization requires greater resolution than that provided by light microscopy. Therefore, researchers seek the much higher resolution provided by electron microscopy. Since 2002, the Cell Sciences Imaging Facility (CSIF) has provided Stanford researchers with a full service EM core lab that provides complete sample preparation, training and support for both transmission and scanning electron microscopy dependent research. The requested sCMOS TEM camera will be used on the EMC?s transmission electron microscope to digitally record TEM images of biomedical samples. It will be used in both high and low electron dose TEM imaging applications and will be used to correct, in real time, the inherent drift of TEM samples. The CSIF has long supported biomedical TEM digital imaging at Stanford and is well positioned to provide ongoing support of this advanced capabilities TEM sCMOS camera. The CSIF?s current TEM camera is a 9 year old, cooled CCD camera that can no longer meet the demanding imaging needs of Stanford?s NIH funded electron microscopy imaging community. This older TEM camera has a slow, limiting frame rate and over the years the scintillator screen and its directly coupled CCD have lost sensitivity and accumulated ?dead? pixels. Additionally, if has a limited field of view, poor signal to noise ratio, and a lower dynamic range compared to newer, state-of-the-art TEM sCMOS cameras. Perhaps most limiting, compared to the requested OneView sCMOS TEM camera, is that the EMC?s current TEM CCD camera lacks drift correction. The requested OneView sCMSO camera will significantly advance the research projects of 15 different users, 14 of which have NIH funded projects and of those, 2 are Noble laureates. These researchers? projects cover a diverse range of biomedically important questions from, for example, the biomechanics of hearing, neural stem cell maintenance and homeostasis, ventricular hypertrophy and failure, to the mechanisms of viral infection and olfaction in sensory neurons, as well as the development of biomedically useful nanoprobes and super-resolution cryogenic correlative light and electron microscopy.
Relevance: The NIH funded biomedical research projects supported by this proposal investigate, in a variety of model organisms, tissues and in vitro systems, diverse aspects of human health and disease, ranging from cancer and viral pathogenesis to understanding the molecular basis of neurodegeneration and sensory biology as well as the development of medically relevant nanoprobes and cryo-imaging techniques. These studies use high- resolution, transmission electron microscopy (TEM) imaging to visualize cell and tissue ultrastructural as well as isolated and assembled proteins, biomolecules and biomaterials. These TEM images need to be recorded digitally and require the requested, state-of-the-art, Gatan Inc., OneView scintillator-coupled sCMOS TEM camera with large field of view, high 16- bit dynamic range, fast frame rate and real time drift correction.
