Translate this page to:
Choose Language…
In JoVE (1)
Other Publications (2)
Articles by Gheorghe Salahura in JoVE
JoVE Neuroscience
Detection of Microregional Hypoxia in Mouse Cerebral Cortex by Two-photon Imaging of Endogenous NADH Fluorescence
1Department of Microbiology and Immunology, University of Rochester Medical Center, 2Center for Neural Development and Disease, University of Rochester Medical Center, 3Deptartment of Neurology, Center for Neural Development and Disease, University of Rochester Medical Center
Here we describe a method to directly visualize microregional tissue hypoxia in the mouse cortex in vivo. It is based on concurrent two-photon imaging of nicotinamide adenine dinucleotide (NADH) and the cortical microcirculation. This method is useful for high resolution analysis of tissue oxygen supply.
Other articles by Gheorghe Salahura on PubMed
Large-scale Propagation of Ultrasound in a 3-D Breast Model Based on High-resolution MRI Data
A 40 x 35 x 25-mm(3) specimen of human breast consisting mostly of fat and connective tissue was imaged using a 3-T magnetic resonance scanner. The resolutions in the image plane and in the orthogonal direction were 130 microm and 150 microm, respectively. Initial processing to prepare the data for segmentation consisted of contrast inversion, interpolation, and noise reduction. Noise reduction used a multilevel bidirectional median filter to preserve edges. The volume of data was segmented into regions of fat and connective tissue by using a combination of local and global thresholding. Local thresholding was performed to preserve fine detail, while global thresholding was performed to minimize the interclass variance between voxels classified as background and voxels classified as object. After smoothing the data to avoid aliasing artifacts, the segmented data volume was visualized using isosurfaces. The isosurfaces were enhanced using transparency, lighting, shading, reflectance, and animation. Computations of pulse propagation through the model illustrate its utility for the study of ultrasound aberration. The results show the feasibility of using the described combination of methods to demonstrate tissue morphology in a form that provides insight about the way ultrasound beams are aberrated in three dimensions by tissue.
Mitochondrial Superoxide Flashes: Metabolic Biomarkers of Skeletal Muscle Activity and Disease
Mitochondrial superoxide flashes (mSOFs) are stochastic events of quantal mitochondrial superoxide generation. Here, we used flexor digitorum brevis muscle fibers from transgenic mice with muscle-specific expression of a novel mitochondrial-targeted superoxide biosensor (mt-cpYFP) to characterize mSOF activity in skeletal muscle at rest, following intense activity, and under pathological conditions. Results demonstrate that mSOF activity in muscle depended on electron transport chain and adenine nucleotide translocase functionality, but it was independent of cyclophilin-D-mediated mitochondrial permeability transition pore activity. The diverse spatial dimensions of individual mSOF events were found to reflect a complex underlying morphology of the mitochondrial network, as examined by electron microscopy. Muscle activity regulated mSOF activity in a biphasic manner. Specifically, mSOF frequency was significantly increased following brief tetanic stimulation (18.1 ± 1.6 to 22.3 ± 2.0 flashes/1000 μm²·100 s before and after 5 tetani) and markedly decreased (to 7.7 ± 1.6 flashes/1000 μm²·100 s) following prolonged tetanic stimulation (40 tetani). A significant temperature-dependent increase in mSOF frequency (11.9 ± 0.8 and 19.8 ± 2.6 flashes/1000 μm²·100 s at 23°C and 37°C) was observed in fibers from RYR1(Y522S/WT) mice, a mouse model of malignant hyperthermia and heat-induced hypermetabolism. Together, these results demonstrate that mSOF activity is a highly sensitive biomarker of mitochondrial respiration and the cellular metabolic state of muscle during physiological activity and pathological oxidative stress
