1University/ BHF Centre for Cardiovascular Science, University of Edinburgh, The Queen's Medical Research Institute
Ex vivo analysis of arterial lesions from animal models of cardiovascular disease classically relies on histological and immunohistochemical techniques. These provide 2-dimensional measurements in 3-dimensional lesions. This manuscript describes the generation of arterial lesions for quantitative analysis in 3-dimensions using optical projection tomography.
Published May 26, 2015. Keywords: Medicine, neointima, mouse femoral artery, atherosclerosis, brachiocephalic trunk, optical projection tomography
1Umeå Centre for Molecular Medicine, Umeå University, 2Cell Transplant Center, Diabetes Research Institute, University of Miami,, 3EMBL-CRG Systems Biology Program, Centre for Genomic Regulation, Catalan Institute of Research and Advanced Studies, 4Dept. of Computing Science, Umeå University
We describe the adaptation of optical projection tomography (OPT)1 to imaging in the near infrared spectrum, and the implementation of a number of computational tools. These protocols enable assessments of pancreatic β-cell mass (BCM) in larger specimens, increase the multichannel capacity of the technique and increase the quality of OPT data.
Published January 12, 2013. Keywords: Medicine, Biomedical Engineering, Cellular Biology, Molecular Biology, Biophysics, Pancreas, Islets of Langerhans, Diabetes Mellitus, Imaging, Three-Dimensional, Optical Projection Tomography, Beta-cell Mass, Near Infrared, Computational Processing
1Breakthrough Breast Cancer Research Unit, Institute of Genetics and Molecular Medicine, University of Edinburgh, 2MRC Technology
We have developed a collagen-based in vitro assay which promotes proliferation and invasion from samples of all breast cancer subtypes. Optical Projection Tomography, a three dimensional microscopy technique was utilised to visualise and quantify tumour expansion. This assay may be used to quantify drug response of individual tumour samples.
Published July 29, 2011. Keywords: Medicine, Breast cancer, Optical Projection Tomography, Imaging, Three-dimensional, computer assisted, Tumour microenvironment
1Center for Systems Biology, Harvard Medical School, 2Center for Systems Biology, MGH - Massachusetts General Hospital, 3Institute for Biological and Medical Imaging, Technical University of Munich and Helmholtz Center Munich
We suggest a Born normalized approach for Optical Projection Tomography (BnOPT) that accounts for the absorption properties of imaged samples to obtain accurate and quantitative fluorescence tomographic reconstructions. We use the proposed algorithm to reconstruct the fluorescence molecular probe distribution within small animal organs.
Published June 2, 2009. Keywords: Bioengineering, optical imaging, fluorescence imaging, optical projection tomography, born normalization, molecular imaging, heart imaging
1Electrical Engineering Department, University of California, Los Angeles, 2Bioengineering Department, University of California, Los Angeles, 3California NanoSystems Institute, University of California, Los Angeles
Lensfree optical tomography is a three-dimensional microscopy technique that offers a spatial resolution of <1 μm × <1 μm × <3 μm in x, y and z dimensions, respectively, over a large imaging-volume of 15-100 mm3, which can be particularly useful for integration with lab-on-a-chip platforms.
Published August 16, 2012. Keywords: Bioengineering, Electrical Engineering, Mechanical Engineering, lensfree imaging, lensless imaging, on-chip microscopy, lensfree tomography, 3D microscopy, pixel super-resolution, C. elegans, optical sectioning, lab-on-a-chip
1U.S. Department of Agriculture, 2Department of Viticulture and Enology, University of California - Davis, 3Hawkesbury Institute for the Environment, University of Western Sydney, 4Advanced Light Source, Lawrence Berkeley National Lab, 5Citrus Research & Education Center, University of Florida
High resolution x-ray computed tomography (HRCT) is a non-destructive diagnostic imaging technique that can be used to study the structure and function of plant vasculature in 3D. We demonstrate how HRCT facilitates exploration of xylem networks across a wide range of plant tissues and species.
Published April 5, 2013. Keywords: Plant Biology, Cellular Biology, Molecular Biology, Biophysics, Structural Biology, Physics, Environmental Sciences, Agriculture, botany, environmental effects (biological, animal and plant), plants, radiation effects (biological, animal and plant), CT scans, advanced visualization techniques, xylem networks, plant vascular function, synchrotron, x-ray micro-tomography, ALS 8.3.2, xylem, phloem, tomography, imaging
1Department of Pathology, Center for Cardiovascular Biology, and Institute for Stem Cell and Regenerative Medicine, University of Washington, 2Departments of Bioengineering and Medicine/Cardiology, University of Washington
Visualization of the coronary vessels is critical to advancing our understanding of cardiovascular diseases. Here we describe a method for perfusing murine coronary vasculature with a radiopaque silicone rubber (Microfil), in preparation for micro-Computed Tomography (μCT) imaging.
Published February 10, 2012. Keywords: Medicine, Vascular biology, heart, coronary vessels, mouse, micro Computed Tomography (μCT) imaging, Microfil
JoVE Developmental Biology
1Department of Medical Biophysics, University of Toronto, 2Sunnybrook Research Institute, 3Lunenfeld-Tanenbaum Research Institute, Mount Sinai Hospital, Toronto
Here, we present a protocol to inject ultrasound microbubble contrast agents into living, isolated late-gestation stage murine embryos. This method enables the study of perfusion parameters and of vascular molecular markers within the embryo using contrast-enhanced high-frequency ultrasound imaging.
Published March 4, 2015. Keywords: Developmental Biology, Micro-ultrasound, Molecular imaging, Mouse embryo, Microbubble, Ultrasound contrast agent, Perfusion
1Department of Biochemistry and Biomedical Sciences, McMaster University, 2Thrombosis and Atherosclerosis Research Institute, McMaster University
We describe procedures to quantify and characterize atherosclerotic lesions in mouse models using precision sectioning of the aortic sinus and ascending aorta combined with histochemical and immunohistochemical analysis.
Published December 7, 2013. Keywords: Medicine, atherosclerosis, atherosclerotic lesion, Mouse Model, aortic sinus, tissue preparation and sectioning, Immunohistochemistry
JoVE Developmental Biology
1Birth Defects Research Centre, UCL Institute of Child Health, 2Blizard Institute, Centre for Digestive Diseases, Queen Mary University of London, Barts and The London School of Medicine and Dentistry, 3Department of Clinical Genetics, Erasmus University Medical Center, Rotterdam
Here we report dual labeling of neural crest cells and blood vessels using chickGFP neural tube intraspecies grafting combined with intra-vascular DiI injection. This experimental technique allows us to simultaneously visualize and study development of the NCC-derived (enteric) nervous system and the vascular system, during organogenesis.
Published May 28, 2015. Keywords: Developmental Biology, Intraspecies grafting, chimera, neural tube, vessel painting, carbocyanine dye, vascular network, transgenic GFP chicken, neural crest cells, enteric nervous system
1School of Biological Sciences, Monash University, 2Janelia Farm Research Campus, Howard Hughes Medical Institute
Time-lapse microscopy allows the visualization of developmental processes. Growth or drift of samples during image acquisition reduces the ability to accurately follow and measure cell movements during development. We describe the use of open source image processing software to correct for three dimensional sample drift over time.
Published April 12, 2014. Keywords: Bioengineering, Image Processing, Computer-Assisted, Zebrafish, Microscopy, Confocal, Time-Lapse Imaging, imaging, zebrafish, Confocal, fiji, three-dimensional, four-dimensional, registration
1Huisken Lab, Max Planck Institute of Molecular Cell Biology and Genetics
The development of zebrafish can be followed over days with light sheet microscopy when embryos are embedded in optically clear polymer tubes with low-concentration agarose.
Published February 27, 2014. Keywords: Developmental Biology, zebrafish, Danio rerio, light sheet microscopy, Selective Plane Illumination Microscopy, sample mounting, time lapse microscopy, long-term imaging
1Department of Cell and Developmental Biology, Center for Stem Cell Biology, Vanderbilt University Medical Center, 2Department of Pharmacology, Center for Stem Cell Biology, Vanderbilt University Medical Center, 3Vanderbilt University Medical Center
Isolation of embryonic neural crest from the neural tube facilitates the use of in vitro methods for studying migration, self-renewal, and multipotency of neural crest.
Published June 2, 2012. Keywords: Neuroscience, Developmental Biology, neural crest, explant, cell culture, mouse, embryo
1Center for Systems Biology, Massachusetts General Hospital, 2Institute for Biological and Medical Imaging (IBMI), Technical University of Munich and Helmholtz Center Munich, 3Department of Genetics, Harvard Medical School and Howard Hughes Medical Institute
Mesoscopic fluorescence tomography operates beyond the penetration limits of tissue-sectioning fluorescence microscopy. The technique is based on multi-projection illumination and a photon transport description. We demonstrate in-vivo whole-body 3D visualization of the morphogenesis of GFP-expressing wing imaginal discs in Drosophila melanogaster.
Published August 20, 2009. Keywords: Developmental Biology, fluorescence tomography, mesoscopic imaging, Drosophila, optical imaging, diffusion tomography, scattering