Analytical Imaging

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PROJECT SUMMARY (See instructions): Imaging plays a vital role in the research programs of many AECC members. The Analytical Imaging Facility (AIF) makes routine and complex imaging technologies available to the entire AECC community. Microscope technology ranges from traditional brightfield, to epi-fluorescence, to laser scanning confocal, to high resolution transmission and scanning electron microscopy including cryoEM of macromolecules in this truly comprehensive shared resource. This wide range of imaging modalities allows for the study of animal models of disease. Cells expressing or labeled with fluorescent reporter molecules may be imaged in the whole animal, whole organ, single cell or cell compartments. Live cells may be imaged in real time to monitor physiological processes. Photo-activatable fluorescence molecules may be manipulated by light for dynamic in vivo studies. Confocal microscopes and epi-fluorescence paired with deconvolution enables 3D imaging of fixed material or 4D imaging of live cells. Traditional ultrastructure analysis by transmission electron microscopy is expanded to 3D by electron tomography. The recent installation of a field emission scanning electron microscope (SEM) has expanded the facility's SEM capabilities to include automated large area mapping, 3D image reconstruction, x-ray microanalysis, correlative fluorescence and SEM imaging and imaging frozen hydrated samples. AIF staff assist users in experimental design, data collection, quantitative image analysis and presentation. New users are taught imaging techniques required for their specific research objective, while experienced users may customize any imaging station, utilizing the facility's large inventory of optics and accessories. The facility offers customized full service sample preparation for electron microscopy, to ensure quality control and to offer the widest range of techniques. The AIF staff has the expertise to prepare samples by methods that include chemical fixation, embedding in resin, ultrathin sectioning, immunogold labeling and negative staining. The facility offers a full range of low temperature techniques for electron microscopy including quick freezing by plunge, metal mirror or high pressure freezing. Frozen samples can undergo freeze-substitution, freeze fracture, rotary shadow or cryosectioning and are then imaged in the TEM or SEM at ambient or low temperature. The AIF provides support for the microscopy requirements of the Molecular Cytogenetics and Histopathology Shared Resources. Since the last CCSG review, important new additions of equipment, personnel and services have been made, which have substantially enhanced the imaging capabilities at AECC.