Browsing by Subject "microscopy"

Now showing 1 - 6 of 6
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    Acridine Orange as a Novel Photosensitizer for Photodynamic Therapy in Glioblastoma
    (Elsevier, 2018) Osman, Hany; Elsahy, Deena; Saadatzadeh, M. Reza; Pollok, Karen E.; Yocom, Steven; Hattab, Eyas; Georges, Joseph; Cohen-Gadol, Aaron A.; Neurological Surgery, School of Medicine
    Object Photodynamic therapy is an exciting treatment modality that combines the effects of a chemical agent with the physical energy from light or radiation to result in lysis of cells of interest. Acridine orange is a molecule with fluorescence properties that was demonstrated to possess photosensitizing properties. The objective of this study was to investigate the photodynamic effect of acridine orange on glioblastoma cell viability and growth. Methods Glioblastoma cells (n = 8000 cells/well at 0 hours) were exposed to acridine orange followed by white unfiltered light-emitting diodes (LED) light. Cultures were exposed to either 10 or 30 minutes of light. The cell number per well was determined at 0, 24, 48, and 72 hours after exposure. Results A dramatic cytocidal effect of acridine orange after exposure to as little as 10 minutes of white light was observed. There was almost complete eradication of the glioblastoma cells over a 72-hour period. Although acridine orange or light alone exhibited some effect on cell growth, it was not as pronounced as the combination of acridine orange and light. Conclusions This is the first study to demonstrate the photodynamic effect of acridine orange in glioblastoma cells. This data supports the need for further studies to characterize and evaluate whether this striking cytotoxic effect can be achieved in vivo. The combination of acridine orange and exposure to white unfiltered LED light may have potential future applications in management of glioblastoma.
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    Direction Selectivity in Drosophila Proprioceptors Requires the Mechanosensory Channel Tmc
    (Elsevier, 2019-03-18) He, Liping; Gulyanon, Sarun; Mihovilovic Skanata, Mirna; Karagyozov, Doycho; Heckscher, Ellie S.; Krieg, Michael; Tsechpenakis, Gavriil; Gershow, Marc; Tracey, W. Daniel; Department of Computer and Information sciences, School of Science
    Summary Drosophila Transmembrane channel-like (Tmc) is a protein that functions in larval proprioception. The closely related TMC1 protein is required for mammalian hearing and is a pore-forming subunit of the hair cell mechanotransduction channel. In hair cells, TMC1 is gated by small deflections of microvilli that produce tension on extracellular tip-links that connect adjacent villi. How Tmc might be gated in larval proprioceptors, which are neurons having a morphology that is completely distinct from hair cells, is unknown. Here, we have used high-speed confocal microscopy both to measure displacements of proprioceptive sensory dendrites during larval movement and to optically measure neural activity of the moving proprioceptors. Unexpectedly, the pattern of dendrite deformation for distinct neurons was unique and differed depending on the direction of locomotion: ddaE neuron dendrites were strongly curved by forward locomotion, while the dendrites of ddaD were more strongly deformed by backward locomotion. Furthermore, GCaMP6f calcium signals recorded in the proprioceptive neurons during locomotion indicated tuning to the direction of movement. ddaE showed strong activation during forward locomotion, while ddaD showed responses that were strongest during backward locomotion. Peripheral proprioceptive neurons in animals mutant for Tmc showed a near-complete loss of movement related calcium signals. As the strength of the responses of wild-type animals was correlated with dendrite curvature, we propose that Tmc channels may be activated by membrane curvature in dendrites that are exposed to strain. Our findings begin to explain how distinct cellular systems rely on a common molecular pathway for mechanosensory responses.
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    Four Dimensional Image Registration For Intravital Microscopy
    (IEEE, 2016-06) Fu, Chichen; Gadgil, Neeraj; Tahboub, Khalid K.; Salama, Paul; Dunn, Kenneth W.; Delp, Edward J.; Department of Biochemistry and Molecular Biology, School of Medicine
    Increasingly the behavior of living systems is being evaluated using intravital microscopy since it provides subcellular resolution of biological processes in an intact living organism. Intravital microscopy images are frequently confounded by motion resulting from animal respiration and heartbeat. In this paper we describe an image registration method capable of correcting motion artifacts in three dimensional fluorescence microscopy images collected over time. Our method uses 3D B-Spline non-rigid registration using a coarse-to-fine strategy to register stacks of images collected at different time intervals and 4D rigid registration to register 3D volumes over time. The results show that our proposed method has the ability of correcting global motion artifacts of sample tissues in four dimensional space, thereby revealing the motility of individual cells in the tissue.
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    Functional studies of the kidney of living animals using multicolor 2-photon microscopy
    (2002-09) Dunn, Kenneth W.; Sandoval, Ruben M.; Kelly, Katherine J.; Dagher, Pierre C.; Tanner, George A.; Atkinson, Simon J.; Bacallao, Robert L.; Molitoris, Bruce A.
    Optical microscopy, when applied to living animals, provides a powerful means of studying cell biology in the most physiologically relevant setting. The ability of two-photon microscopy to collect optical sections deep into biological tissues has opened up the field of intravital microscopy to high-resolution studies of the brain, lens, skin, and tumors. Here we present examples of the way in which two-photon microscopy can be applied to intravital studies of kidney physiology. Because the kidney is easily externalized without compromising its function, microscopy can be used to evaluate various aspects of renal function in vivo. These include cell vitality and apoptosis, fluid transport, receptor-mediated endocytosis, blood flow, and leukocyte trafficking. Efficient two-photon excitation of multiple fluorophores permits comparison of multiple probes and simultaneous characterization of multiple parameters and yields spectral information that is crucial to the interpretation of images containing uncharacterized autofluorescence. The studies described here demonstrate the way in which two-photon microscopy can provide a level of resolution previously unattainable in intravital microscopy, enabling kinetic analyses and physiological studies of the organs of living animals with subcellular resolution.
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    In Vivo Microscopy in Neurosurgical Oncology
    (Elsevier, 2018) Osman, Hany; Georges, Joseph; Elsahy, Deena; Hattab, Eyas; Yocom, Steven; Cohen-Gadol, Aaron A.; Neurological Surgery, School of Medicine
    Intraoperative neurosurgical histopathologic diagnoses rely on evaluation of rapid tissue preparations such as frozen sections and smears with conventional light microscopy. Though useful, these techniques are time consuming and therefore unable to provide real-time intraoperative feedback. In vivo molecular imaging techniques are emerging as novel methods for generating real-time diagnostic histopathologic images of tumors and their surrounding tissues. These imaging techniques rely on contrast generated by exogenous fluorescent dyes, autofluorescence of endogenous molecules, fluorescence decay of excited molecules, or light scattering. Large molecular imaging instruments are being miniaturized for clinical in vivo use. This review discusses pertinent imaging systems that have been developed for neurosurgical use and imaging techniques currently under NADPH development for neurosurgical molecular imaging.
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    Three Dimensional Fluorescence Microscopy Image Synthesis and Segmentation
    (IEEE, 2018-06) Fu, Chichen; Lee, Soonam; Ho, David Joon; Han, Shuo; Salama, Paul; Dunn, Kenneth W.; Delp, Edward J.; Electrical and Computer Engineering, School of Engineering and Technology
    Advances in fluorescence microscopy enable acquisition of 3D image volumes with better image quality and deeper penetration into tissue. Segmentation is a required step to characterize and analyze biological structures in the images and recent 3D segmentation using deep learning has achieved promising results. One issue is that deep learning techniques require a large set of groundtruth data which is impractical to annotate manually for large 3D microscopy volumes. This paper describes a 3D deep learning nuclei segmentation method using synthetic 3D volumes for training. A set of synthetic volumes and the corresponding groundtruth are generated using spatially constrained cycle-consistent adversarial networks. Segmentation results demonstrate that our proposed method is capable of segmenting nuclei successfully for various data sets.