Browsing by Subject "intravital microscopy"

Now showing 1 - 4 of 4
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    Intravital imaging of the kidney
    (Elsevier, 2017-09-01) Hato, Takashi; Winfree, Seth; Dagher, Pierre C.; Medicine, School of Medicine
    Two-photon intravital microscopy is a powerful tool that allows the examination of dynamic cellular processes in the live animal with unprecedented resolution. Indeed, it offers the ability to address unique biological questions that may not be solved by other means. While two-photon intravital microscopy has been successfully applied to study many organs, the kidney presents its own unique challenges that need to be overcome in order to optimize and validate imaging data. For kidney imaging, the complexity of renal architecture and salient autofluorescence merit special considerations as these elements directly impact image acquisition and data interpretation. Here, using illustrative cases, we provide practical guides and discuss issues that may arise during two-photon live imaging of the rodent kidney.
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    Intravital microscopy of biosensor activities and intrinsic metabolic states
    (Elsevier, 2017-09-01) Winfree, Seth; Hato, Takashi; Day, Richard N.; Department of Cellular & Integrative Physiology, Indiana University School of Medicine
    Intravital microscopy (IVM) is an imaging tool that is capable of detecting subcellular signaling or metabolic events as they occur in tissues in the living animal. Imaging in highly scattering biological tissues, however, is challenging because of the attenuation of signal in images acquired at increasing depths. Depth-dependent signal attenuation is the major impediment to IVM, limiting the depth from which significant data can be obtained. Therefore, making quantitative measurements by IVM requires methods that use internal calibration, or alternatively, a completely different way of evaluating the signals. Here, we describe how ratiometric imaging of genetically encoded biosensor probes can be used to make quantitative measurements of changes in the activity of cell signaling pathways. Then, we describe how fluorescence lifetime imaging can be used for label-free measurements of the metabolic states of cells within the living animal.
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    Surgical Preparation Of Rats And Mice For Intravital Microscopic Imaging Of Abdominal Organs
    (Elsevier, 2017) Rhodes, George J.; Department of Medicine, School of Medicine
    Intravital microscopy is a powerful research tool that can provide insight into cellular and subcellular events that take place in organs in the body. However, meaningful results can only be obtained from animals whose physiology is preserved during the process of microscopy. Here I discuss the importance of preserving the overall state of health of the animal, methods of anesthesia, surgical techniques for intravital microscopy of various abdominal organs, methods to maintain and monitor the physiology of the animal during microscopy and associated peri- and post-operative recovery considerations.
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    Using quantitative intravital multiphoton microscopy to dissect hepatic transport in rats
    (Elsevier, 2017) Dunn, Kenneth W.; Ryan, Jennifer C.; Department of Medicine, IU School of Medicine
    Hepatic solute transport is a complex process whose disruption is associated with liver disease and drug-induced liver injury. Intravital multiphoton fluorescence excitation microscopy provides the spatial and temporal resolution necessary to characterize hepatic transport at the level of individual hepatocytes in vivo and thus to identify the mechanisms and cellular consequences of cholestasis. Here we present an overview of the use of fluorescence microscopy for studies of hepatic transport in living animals, and describe how we have combined methods of intravital microscopy and digital image analysis to dissect the effects of drugs and pathological conditions on the function of hepatic transporters in vivo.