Browsing by Subject "In vivo imaging"

Now showing 1 - 5 of 5
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    Combining Intravital Fluorescent Microscopy (IVFM) with Genetic Models to Study Engraftment Dynamics of Hematopoietic Cells to Bone Marrow Niches
    (Journal of Visualized Experiments, 2017-03-21) Wang, Lin; Kamocka, Malgorzata M.; Zollman, Amy; Carlesso, Nadia; Pediatrics, School of Medicine
    Increasing evidence indicates that normal hematopoiesis is regulated by distinct microenvironmental cues in the BM, which include specialized cellular niches modulating critical hematopoietic stem cell (HSC) functions1,2. Indeed, a more detailed picture of the hematopoietic microenvironment is now emerging, in which the endosteal and the endothelial niches form functional units for the regulation of normal HSC and their progeny3,4,5. New studies have revealed the importance of perivascular cells, adipocytes and neuronal cells in maintaining and regulating HSC function6,7,8. Furthermore, there is evidence that cells from different lineages, i.e. myeloid and lymphoid cells, home and reside in specific niches within the BM microenvironment. However, a complete mapping of the BM microenvironment and its occupants is still in progress. Transgenic mouse strains expressing lineage specific fluorescent markers or mice genetically engineered to lack selected molecules in specific cells of the BM niche are now available. Knock-out and lineage tracking models, in combination with transplantation approaches, provide the opportunity to refine the knowledge on the role of specific "niche" cells for defined hematopoietic populations, such as HSC, B-cells, T-cells, myeloid cells and erythroid cells. This strategy can be further potentiated by merging the use of two-photon microscopy of the calvarium. By providing in vivo high resolution imaging and 3-D rendering of the BM calvarium, we can now determine precisely the location where specific hematopoietic subsets home in the BM and evaluate the kinetics of their expansion over time. Here, Lys-GFP transgenic mice (marking myeloid cells)9 and RBPJ knock-out mice (lacking canonical Notch signaling)10 are used in combination with IVFM to determine the engraftment of myeloid cells to a Notch defective BM microenvironment.
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    Convolutional neural network denoising in fluorescence lifetime imaging microscopy (FLIM)
    (SPIE, 2021) Mannam, Varun; Zhang, Yide; Yuan, Xiaotong; Hato, Takashi; Dagher, Pierre C.; Nichols, Evan L.; Smith, Cody J.; Dunn, Kenneth W.; Howard, Scott; Medicine, School of Medicine
    Fluorescence lifetime imaging microscopy (FLIM) systems are limited by their slow processing speed, low signal- to-noise ratio (SNR), and expensive and challenging hardware setups. In this work, we demonstrate applying a denoising convolutional network to improve FLIM SNR. The network will integrated with an instant FLIM system with fast data acquisition based on analog signal processing, high SNR using high-efficiency pulse-modulation, and cost-effective implementation utilizing off-the-shelf radio-frequency components. Our instant FLIM system simultaneously provides the intensity, lifetime, and phasor plots in vivo and ex vivo. By integrating image de- noising using the trained deep learning model on the FLIM data, provide accurate FLIM phasor measurements are obtained. The enhanced phasor is then passed through the K-means clustering segmentation method, an unbiased and unsupervised machine learning technique to separate different fluorophores accurately. Our experimental in vivo mouse kidney results indicate that introducing the deep learning image denoising model before the segmentation effectively removes the noise in the phasor compared to existing methods and provides clearer segments. Hence, the proposed deep learning-based workflow provides fast and accurate automatic segmentation of fluorescence images using instant FLIM. The denoising operation is effective for the segmentation if the FLIM measurements are noisy. The clustering can effectively enhance the detection of biological structures of interest in biomedical imaging applications.
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    Editorial: Pancreas Imaging Across the Spectrum
    (Frontiers Media, 2021-01-03) Linnemann, Amelia K.; Poitout, Vincent; Rutter, Guy A.; Pediatrics, School of Medicine
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    Fast assessment of lipid content in arteries in vivo by intravascular photoacoustic tomography
    (Nature Publishing Group, 2018-02-05) Cao, Yingchun; Kole, Ayeeshik; Hui, Jie; Zhang, Yi; Mai, Jieying; Alloosh, Mouhamad; Sturek, Michael; Cheng, Ji-Xin; Cellular and Integrative Physiology, School of Medicine
    Intravascular photoacoustic tomography is an emerging technology for mapping lipid deposition within an arterial wall for the investigation of the vulnerability of atherosclerotic plaques to rupture. By converting localized laser absorption in lipid-rich biological tissue into ultrasonic waves through thermoelastic expansion, intravascular photoacoustic tomography is uniquely capable of imaging the entire arterial wall with chemical selectivity and depth resolution. However, technical challenges, including an imaging catheter with sufficient sensitivity and depth and a functional sheath material without significant signal attenuation and artifact generation for both photoacoustics and ultrasound, have prevented in vivo application of intravascular photoacoustic imaging for clinical translation. Here, we present a highly sensitive quasi-collinear dual-mode photoacoustic/ultrasound catheter with elaborately selected sheath material, and demonstrated the performance of our intravascular photoacoustic tomography system by in vivo imaging of lipid distribution in rabbit aortas under clinically relevant conditions at imaging speeds up to 16 frames per second. Ex vivo evaluation of fresh human coronary arteries further confirmed the performance of our imaging system for accurate lipid localization and quantification of the entire arterial wall, indicating its clinical significance and translational capability.
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    Imaging Neural Activity in the Primary Somatosensory Cortex Using Thy1-GCaMP6s Transgenic Mice
    (Journal of Visualized Experiments (JoVE), 2019-01-07) Lin, Xiaojing; Zhao, Tingbao; Xiong, Wenhui; Wen, Shaonan; Jin, Xiaoming; Xu, Xiao-Ming; Neurological Surgery, School of Medicine
    The mammalian brain exhibits marked symmetry across the sagittal plane. However, detailed description of neural dynamics in symmetric brain regions in adult mammalian animals remains elusive. In this study, we describe an experimental procedure for measuring calcium dynamics through dual optical windows above bilateral primary somatosensory corticies (S1) in Thy1-GCaMP6s transgenic mice using 2-photon (2P) microscopy. This method enables recordings and quantifications of neural activity in bilateral mouse brain regions one at a time in the same experiment for a prolonged period in vivo. Key aspects of this method, which can be completed within an hour, include minimally invasive surgery procedures for creating dual optical windows, and the use of 2P imaging. Although we only demonstrate the technique in the S1 area, the method can be applied to other regions of the living brain facilitating the elucidation of structural and functional complexities of brain neural networks.