1Experimental Radiology, Institute of Diagnostic and Interventional Radiology I, Jena University Hospital, 2Department of Pharmaceutical Technology, Friedrich-Schiller-University Jena, 3Center for Electron Microscopy, Jena University Hospital
The use of fluorophores for in vivo imaging can be greatly limited by opsonization, rapid clearance, low detection sensitivity and cytotoxic effects on the host. Encapsulation of fluorophores in liposomes by film hydration and extrusion leads to fluorescence quenching and protection which enables in vivo imaging with high detection sensitivity.
Published January 5, 2015. Keywords: Bioengineering, Drug-delivery, Liposomes, Fluorochromes, Fluorescence-quenching, Optical imaging, Inflammation
1Functional and Applied Biomechanics Section, Rehabilitation Medicine Department, Clinical Center, National Institutes of Health
Monitoring brain activity during upright motor tasks is of great value when investigating the neural source of movement disorders. Here, we demonstrate a protocol that combines functional near infrared spectroscopy with continuous monitoring of muscle and kinematic activity during 4 types of motor tasks.
Published December 5, 2014. Keywords: Behavior, functional near infrared spectroscopy, fNIRS, brain activity, gait, motor tasks, cerebral palsy, coordination
1School of Biomedical Engineering, Science and Health Systems, Drexel University, 2College of Nursing and Health Professions, Drexel University
MazeSuite is a complete toolset to prepare, present and analyze navigational and spatial experiments. Functional near-infrared spectroscopy (fNIR) is an optical brain imaging technique that enables noninvasive and portable monitoring of cerebral blood oxygenation changes. This paper summarizes collective use of MazeSuite and fNIR within a cognitive processing learning paradigm.
Published October 8, 2011. Keywords: Neuroscience, Cognitive, optical, brain, imaging, functional near-infrared spectroscopy, fNIR, spatial, navigation, software
1Department of Surgery, University Medical Center Groningen, 2Helmholtz Zentrum Munich, Technical University of Munich
Near-infrared fluorescence (NIRF) imaging may improve therapeutic outcome of breast cancer surgery by enabling intraoperative tumor localization and evaluation of surgical margin status. Using tissue-simulating breast phantoms containing fluorescent tumor-simulating inclusions, potential clinical applications of NIRF imaging in breast cancer patients can be assessed for standardization and training purposes.
Published September 19, 2014. Keywords: Medicine, Breast cancer, tissue-simulating phantoms, NIRF imaging, tumor-simulating inclusions, fluorescence, intraoperative imaging
1Discipline of Physiology, University of Sydney, 2Discipline of Biomedical Science, University of Sydney
Mitochondrial dysfunction is a hallmark of cellular senescence. This paper uses non-invasive near-infrared (NIr) treatment to improve mitochondrial function in the aging mouse vestibular sensory epithelium.
Published March 14, 2015. Keywords: Molecular Biology, Vestibular, mitochondria, infrared, neuroprotection, aging, oxidative stress
1Cardiovascular Research Center and Cardiology Division, Massachusetts General Hospital, Harvard Medical School, 2Institute for Biological and Medical Imaging, Helmholtz Zentrum München und Technische Universität München, 3Department of Electrical and Computer Engineering, Northeastern University
We detail a new near-infrared fluorescence (NIRF) catheter for 2-dimensional intravascular molecular imaging of plaque biology in vivo. The NIRF catheter can visualize key biological processes such as inflammation by reporting on the presence of plaque-avid activatable and targeted NIR fluorochromes. The catheter utilizes clinical engineering and power requirements and is targeted for application in human coronary arteries. The following research study describes a multimodal imaging strategy that utilizes a novel in vivo intravascular NIRF catheter to image and quantify inflammatory plaque in proteolytically active inflamed rabbit atheromata.
Published August 4, 2011. Keywords: Medicine, Atherosclerosis, inflammation, imaging, near infrared fluorescence, plaque, intravascular, catheter
1Experimental and Regenerative Neurosciences, School of Animal Biology, The University of Western Australia, 2School of Animal Biology and The Oceans Institute, The University of Western Australia, 3Experimental and Regenerative Neurosciences, School of Anatomy, Physiology and Human Biology, The University of Western Australia
Non-coherent Xenon light was passed through narrow-band interference and neutral density filters to deliver light of varying wavelength and intensity to cultured cells. This protocol was used to assess the effects of red/near-infrared light therapy on production of reactive species in vitro: no effects were observed using the tested parameters.
Published March 21, 2015. Keywords: Engineering, Red light therapy, reactive oxygen species, oxidative stress, photobiomodulation, optimization, irradiation
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
1Department of Psychology, University of Michigan, Ann Arbor, 2Department of Psychology, University of Toronto Scarborough
Here we describe a data collection and data analysis method for functional Near Infrared Spectroscopy (fNIRS), a novel non-invasive brain imaging system used in cognitive neuroscience, particularly in studying child brain development. This method provides a universal standard of data acquisition and analysis vital to data interpretation and scientific discovery.
Published July 28, 2009. Keywords: Neuroscience, infant, child, Near Infrared Spectroscopy, fNIRS, optical tomography, cognitive neuroscience, psychology, brain, developmental cognitive neuroscience, analysis
1Division of Pharmaceutical Sciences, Mylan School of Pharmacy, Duquesne University
Therapeutic compounds are often first examined in vitro with viability assays. Blind cell counts by a human observer can be highly sensitive to small changes in cell number but do not assess function. Computerized viability assays, as described here, can assess both structure and function in an objective manner.
Published January 20, 2014. Keywords: Cellular Biology, In-cell Western, DRAQ5, Sapphire, Cell Titer Glo, ATP, primary cortical neurons, toxicity, protection, N-acetyl cysteine, hormesis
1Biotactical Engineering, Faculty of Engineering and Industrial Science, Swinburne University of Technology, 2Department of Otolaryngology, The University of Melbourne
Infrared nerve stimulation has been proposed as an alternative to electrical stimulation in a range of nerve types, including those associated with the auditory system. This protocol describes a patch clamp method for studying the mechanism of infrared nerve stimulation in a culture of primary auditory neurons.
Published July 31, 2013. Keywords: Neuroscience, Biomedical Engineering, Neurobiology, Molecular Biology, Cellular Biology, Physiology, Primary Cell Culture, Biophysics, Electrophysiology, fiber optics, infrared neural stimulation, patch clamp, in vitro models, spiral ganglion neurons, neurons, patch clamp recordings, cell culture
1Diller Cancer Research Building, University of California, San Francisco
This protocol explores the latest advancements in performing Western blot analyses. These novel modifications employ a Bis-Tris gel system with a 35 min electrophoresis run time, a 7 min dry blotting transfer system, and infrared fluorescent protein detection and imaging that generates higher resolution, quality, sensitivity, and improved accuracy of Western data.
Published February 5, 2014. Keywords: Basic Protocol, Western blot, Bis-Tris, electrophoresis, dry blotting, protein transfer, infrared, Fluorescence, quantification, Antibody, Protein
1Department of Diagnostic and Interventional Radiology, University Medical Center, Hamburg, 2Institute of Immunology, University Medical Center, Hamburg, 3University Cancer Center Hamburg, University Medical Center, Hamburg, 4Department of Oncology and Hematology, University Medical Center, Hamburg
This protocol outlines the steps required to perform ex vivo validation of in vivo near-infrared fluorescence xenograft imaging experiments in mice using fluorophore labelled nanobodies and conventional antibodies.
Published April 6, 2015. Keywords: Medicine, Nanobody, antibody, VHH, fluorescence imaging, molecular imaging, xenograft, animal model
1Department of Chemistry, The University of Georgia
The preparation and exfoliation of CaCuSi4O10 and BaCuSi4O10 are described. Upon stirring in hot water, CaCuSi4O10 spontaneously exfoliates into monolayers, whereas BaCuSi4O10 requires ultrasonication in organic solvents. Near infrared (NIR) imaging illustrates the NIR emission properties of these materials, and aqueous dispersions of these nanomaterials are useful for solution processing.
Published April 24, 2014. Keywords: Chemistry, Nanosheets, Egyptian Blue, Han Blue, Pigment, Near Infrared, Luminescence, Exfoliation, Delamination, Two-Dimensional, Ink, Colloidal Dispersion
JoVE Immunology and Infection
1Center for Molecular Imaging (CMI), University of Texas Health Science Center-Houston
Recently developed imaging techniques using near-infrared fluorescence (NIRF) may help elucidate the role the lymphatic system plays in cancer metastasis, immune response, wound repair, and other lymphatic-associated diseases.
Published March 8, 2013. Keywords: Immunology, Medicine, Anatomy, Physiology, Molecular Biology, Biomedical Engineering, Cancer Biology, Optical imaging, lymphatic imaging, mouse imaging, non-invasive imaging, near-infrared fluorescence, vasculature, circulatory system, lymphatic system, lymph, dermis, injection, imaging, mouse, animal model
1Department of Physics, University of Ottawa, 2Department of Cellular and Molecular Medicine, University of Ottawa, 3Centre for Neural Dynamics, University of Ottawa
We describe a set of techniques for studying spontaneous behavior of freely swimming weakly electric fish over an extended period of time, by synchronously measuring the animal's electric organ discharge timing, body position and posture both accurately and reliably in a specially designed aquarium tank inside a sensory isolation chamber.
Published March 6, 2014. Keywords: Neuroscience, animal tracking, weakly electric fish, electric organ discharge, underwater infrared imaging, automated image tracking, sensory isolation chamber, exploratory behavior
1Department of Biomedical Engineering, The Ohio State University, 2Department of Biomedical Informatics, The Ohio State University, 3Comprehensive Wound Center, The Ohio State University, 4Department of Surgery, The Ohio State University
A dual-mode imaging system was developed for non-contact assessment of cutaneous tissue oxygenation and vascular function.
Published December 8, 2010. Keywords: Medicine, Dual-mode, multispectral imaging, infrared imaging, cutaneous tissue oxygenation, vascular function, co-registration, wound healing
1ARC Centre of Excellence for Electromaterials Science (ACES), Intelligent Polymer Research Institute (IPRI), The University of Wollongong, 2School of Chemistry, The University of Sydney, 3School of Chemistry, The University of New South Wales
An integrated device, incorporating a dye-sensitized solar cell and triplet-triplet annihilation up-conversion unit was produced, affording enhanced light harvesting, from a wider section of the solar spectrum. Under modest irradiation levels a significantly enhanced response to low energy photons was demonstrated, yielding a record figure of merit for dye-sensitized solar cells.
Published September 12, 2014. Keywords: Physics, Third generation photovoltaics; upconversion; organic electronics; device architecture; porphyrins; photovoltaic testing
1Athinoula A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital, Harvard Medical School, 2Lab. PALM, Université de Caen Basse-Normandie, 3Fetal-Neonatal Neuroimaging and Developmental Science Center, Boston Children's Hospital, Harvard Medical School, 4ISS, INC.
We combined frequency-domain near-infrared spectroscopy measures of cerebral hemoglobin oxygenation with diffuse correlation spectroscopy measures of cerebral blood flow index to estimate an index of oxygen metabolism. We tested the utility of this measure as a bedside screening tool to evaluate the health and development of the newborn brain.
Published March 14, 2013. Keywords: Medicine, Developmental Biology, Neurobiology, Neuroscience, Biomedical Engineering, Anatomy, Physiology, Near infrared spectroscopy, diffuse correlation spectroscopy, cerebral hemodynamic, cerebral metabolism, brain injury screening, brain health, brain development, newborns, neonates, imaging, clinical techniques
1Department of Pathology and Immunology, Faculty of Medicine, University of Geneva, 2Physics Department, GAP-Biophotonics, University of Geneva, 3Laboratoire d'Optique Biomédicale (LOB), Faculté des Sciences et Techniques de l'Ingénieur, École Polytechnique Fédérale de Lausanne, 4Department of Clinical Medicine, School of Medicine, Trinity College Dublin, 5School of Medicine and CRANN, Trinity College Dublin, 6Nikon AG Instruments
Protocol details are provided for in vitro labeling human embryonic stem cells with second harmonic generating nanoparticles. Methodologies for hESC investigation by multi-photon microscopy and their differentiation into cardiac clusters are also presented.
Published May 1, 2014. Keywords: Bioengineering, multi-photon imaging, human embryonic stem cells (ESC), nanoparticles, embryoid bodies (EBs), cardiomyocyte differentiation, cardiac contraction, air-liquid cultures
1Department of Industrial and Physical Pharmacy, Purdue University
Here, we present detailed protocols for solid-state amide hydrogen/deuterium exchange mass spectrometry (ssHDX-MS) and solid-state photolytic labeling mass spectrometry (ssPL-MS) for proteins in solid powders. The methods provide high-resolution information on protein conformation and interactions in the amorphous solid-state, which may be useful in formulation design.
Published April 14, 2015. Keywords: Chemistry, Amide hydrogen/deuterium exchange, photolytic labeling, mass spectrometry, lyophilized formulations, photo-leucine, solid-state, protein structure, protein conformation, protein dynamics, secondary structure, protein stability, excipients
1InnerEarLab, Department of Otolaryngology, University Medical Center Goettingen, 2Bernstein Focus for Neurotechnology, University of Goettingen, 3Auditory Systems Physiology Group, Department of Otolaryngology, University Medical Center Goettingen, 4Center for Nanoscale Microscopy and Molecular Physiology of the Brain, University of Goettingen, 5Department of Chemical, Electronic, and Biomedical Engineering, University of Guanajuato
Cochlear implants (CIs) enable hearing by direct electrical stimulation of the auditory nerve. However, poor frequency and intensity resolution limits the quality of hearing with CIs. Here we describe optogenetic stimulation of the auditory nerve in mice as an alternative strategy for auditory research and developing future CIs.
Published October 8, 2014. Keywords: Neuroscience, hearing, cochlear implant, optogenetics, channelrhodopsin, optical stimulation, deafness
1Department of Physics and Astronomy, The University of Texas at San Antonio, 2Centro de Investigaciones en Optica A. C., 3Department of Biology and Neurosciences Institute, The University of Texas at San Antonio
We synthesized star shaped gold nanostars using a silver seed mediated growth method. The diameter of the nanostars ranges from 200 to 300 nm and the number of tips vary from 7 to 10. The nanoparticles have a broad surface plasmon resonance mode centered in the near infrared.
Published January 15, 2012. Keywords: Bioengineering, thermal ablation, surface plasmon resonance, nanoparticle, nanotechnology, silver seeds
1Laboratoire Interdisciplinaire Carnot de Bourgogne CNRS-UMR 6303, Université de Bourgogne, 2Department of Optics and Optical Engineering, University of Science and Technology of China, 3CEMES, CNRS-UPR 8011
Silver nanowires can simultaneously transport electrons and optical information in the form of surface plasmons. A procedure is described here to realize such a shared circuitry and the limitations at propagating both information carriers are evaluated.
Published December 11, 2013. Keywords: Physics, light transmission, optical waveguides, photonics, plasma oscillations, plasma waves, electron motion in conductors, nanofabrication, Information Transport, plasmonics, Silver Nanowires, Leakage radiation microscopy, Electromigration
1LENS - European Laboratory for Non-linear Spectroscopy, University of Florence, 2Chemistry Research Laboratory, University of Oxford, 3Department of Biology, University of Florence, 4Department of Physics and Astronomy, University of Florence, 5National Institute of Optics-National Research Council, Italy, 6International Center of Computational Neurophotonics
Here we describe the instrumentation and methods for detecting single fluorescently-labeled protein molecules interacting with a single DNA molecule suspended between two optically trapped microspheres.
Published August 27, 2014. Keywords: Bioengineering, Single molecule biophysics, Optical tweezers, fluorescence microscopy, DNA binding proteins, lactose repressor, microfluidics
1Plastic and Reconstructive Surgery, Johns Hopkins University School of Medicine
Vascularized Composite Allotransplantations (VCA) have become a clinical reality. However, broad clinical application of VCA is limited by chronic multi-drug immunosuppression. The authors present a reliable and reproducible large animal model to translate novel immunomodulatory strategies that can minimize or potentially eliminate the need of immunosuppression in VCA.
Published October 14, 2013. Keywords: Medicine, Upper Extremity, Swine, Microsurgery, Tissue Transplantation, Transplantation Immunology, Surgical Procedures, Operative, Vascularized Composite Allografts, reconstructive transplantation, translational research, swine, hind limb allotransplantation, bone marrow, osteomyocutaneous, microvascular anastomosis, immunomodulation
JoVE Application Notes
Quantitative measurement of cleaved PARP-1 in fixed adherent or suspension cells by high-throughput In-Cell ELISA for using infra-red Li-Cor imaging system.
Published June 14, 2012. Keywords: Advertisement, In-Cell ELISA, Odyssey, apoptosis marker, PARP-1, ICE, quantitation, immunoassay, Genetics
1School of Engineering, University of Glasgow
This protocol outlines the simulation, fabrication and characterization of THz metamaterial absorbers. Such absorbers, when coupled with an appropriate sensor, have applications in THz imaging and spectroscopy.
Published December 27, 2012. Keywords: Materials Science, Physics, Engineering, Metamaterial, terahertz, sensing, fabrication, clean room, simulation, FTIR, spectroscopy
1Department of Biomedical Engineering, University of Texas at Austin, 2Department of Imaging Physics, University of Texas M.D. Anderson Cancer Center
Here, we describe a protocol for synthesis of magneto-plasmonic nanoparticles with a strong magnetic moment and a strong near-infrared (NIR) absorbance. The protocol also includes antibody conjugation to the nanoparticles through the Fc moiety for various biomedical applications which require molecular specific targeting.
Published August 22, 2014. Keywords: Chemistry, nanoparticles, plasmonic, magnetic, nanocomposites, magnetic trapping, circulating tumor cells, dark-field imaging
1Department of Biomedical Engineering, Emory University, 2Department of Chemistry, Georgia Institute of Technology
We describe the preparation of colloidal quantum dots with minimized hydrodynamic size for single-molecule fluorescence imaging. Compared to conventional quantum dots, these nanoparticles are similar in size to globular proteins and are optimized for single-molecule brightness, stability against photodegradation, and resistance to nonspecific binding to proteins and cells.
Published October 9, 2012. Keywords: Physics, Biomedical Engineering, Chemistry, Nanotechnology, Nanoparticle, nanocrystal, synthesis, fluorescence, microscopy, imaging, conjugation, dynamics, intracellular, receptor
1Biomedical Engineering, Laboratory for Fluorescence Dynamics, University of California, Irvine
In this video protocol we track - at high speed and in three dimensions - fluorescently labeled lysosomes within living cells, using the orbital tracking method in a modified two-photon microscope.
Published October 1, 2014. Keywords: Bioengineering, fluorescence, single particle tracking, laser scanning microscope, two-photon, vesicle transport, live-cell imaging, optics
1Department of Molecular Genetics, The Ohio State University
Here, a novel quantitative fluorescence assay is developed to measure changes in the level of a protein specifically at centrosomes by normalizing that protein’s fluorescence intensity to that of an appropriate internal standard.
Published December 20, 2014. Keywords: Cellular Biology, Centrosome assembly, cell cycle, centrosomal degradation, quantitative fluorescence microscopy, normalization, VDAC3, BrdU pulse-chase
1Department of Physics, Bowling Green State University, 2The Center for Photochemical Sciences, Bowling Green State University, 3Department of Chemistry, Bowling Green State University
A general strategy for the development of charge-separating semiconductor nanocrystal composites deployable for solar energy production is presented. We show that assembly of donor-acceptor nanocrystal domains in a single nanoparticle geometry gives rise to a photocatalytic function, while bulk-heterojunctions of donor-acceptor nanocrystal films can be used for photovoltaic energy conversion.
Published August 23, 2012. Keywords: Physics, Materials Science, Chemical Engineering, Chemistry, Electrical Engineering, Photovoltaics, nanorods, dye-sensitized, solids, titanium dioxide, photocatalysis, quantum dots
1Department of Bioengineering, University of Utah, 2Department of Radiology and Radiological Science, Johns Hopkins University School of Medicine, 3Institute for NanoBiotechnology, Johns Hopkins University
This procedure demonstrates in vivo near IR fluorescence imaging of collagen remodeling activities in mice as well as ex vivo staining of collagens in tissue sections using caged collagen mimetic peptides that can be photo-triggered to hybridize with denatured collagen strands.
Published January 31, 2014. Keywords: Bioengineering, collagen remodeling, triple helix, near infrared fluorescence, bioimaging, tissue staining
1School of Physics & Astronomy, University of St Andrews
Use of photonic crystal slow light waveguides and cavities has been widely adopted by the photonics community in many differing applications. Therefore fabrication and characterization of these devices are of great interest. This paper outlines our fabrication technique and two optical characterization methods, namely: interferometric (waveguides) and resonant scattering (cavities).
Published November 30, 2012. Keywords: Physics, Optics and Photonics, Astronomy, light scattering, light transmission, optical waveguides, photonics, photonic crystals, Slow-light, Cavities, Waveguides, Silicon, SOI, Fabrication, Characterization
1Institute for Genomic Biology, University of Illinois at Urbana-Champaign, 2Department of Geology, University of Illinois at Urbana-Champaign, 3Department of Microbiology, University of Illinois at Urbana-Champaign
An integrated suite of imaging techniques has been applied to determine polyp morphology and tissue structure in the Caribbean corals Montastraeaannularis and M. faveolata. Fluorescence, serial block face, and two-photon confocal laser scanning microscopy have identified lobate structure, polyp walls, and estimated chromatophore and zooxanthellae densities and distributions.
Published September 5, 2014. Keywords: Environmental Sciences, Serial block face imaging, two-photon fluorescence microscopy, Montastraea annularis, Montastraea faveolata, 3D coral tissue morphology and structure, zooxanthellae, chromatophore, autofluorescence, light harvesting optimization, environmental change
1Thayer School of Engineering, Dartmouth College, 2Department of Physics and Astronomy, Dartmouth College, 3Darmouth Medical School, Dartmouth College, 4School of Computer Science, University of Birmingham
Diffuse fluorescence tomography offers a relatively low-cost and potentially high-throughout approach to preclinical in vivo tumor imaging. The methodology of optical data collection, calibration, and image reconstruction is presented for a computed tomography-guided non-contact time-domain system using fluorescent targeting of the tumor biomarker epidermal growth factor receptor in a mouse glioma model.
Published July 17, 2012. Keywords: Cancer Biology, Medicine, Physics, Molecular Biology, fluorescence, glioma, light transport, tomography, CT, molecular imaging, epidermal growth factor receptor, biomarker
1Laboratoire Matière et Systèmes Complexes (MSC), CNRS/Université Paris Diderot, 2ImagoSeine BioImaging Core Facility, Institut Jacques Monod, CNRS/Université Paris Diderot, 3Laboratoire d'Immunopathologie et Chimie Thérapeutique, CNRS/Institut de Biologie Moléculaire et Cellulaire
In this study, the main purpose was to monitor the cellular uptake and eventual exocytosis of carbon nanotubes (CNTs). From this perspective, we proposed here a unique method using multispectral imaging flow cytometry allowing a quantification and localization of CNTs in a statistically relevant number of cells.
Published December 12, 2013. Keywords: Bioengineering, bioengineering, imaging flow cytometry, Carbon Nanotubes, bio-nano-interactions, cellular uptake, cell trafficking
1Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University
In this paper, we describe a useful method to study ligand-gated ion channel function in neurons of acutely isolated brain slices. This method involves the use of a drug-filled micropipette for local application of drugs to neurons recorded using standard patch clamp techniques.
Published October 29, 2012. Keywords: Neuroscience, Nicotinic, acetylcholine, neurotransmitter, neuron, patch clamp, brain slice, picospritzer
1Department of Biomedical Engineering, Case Western Reserve University, 2Department of Biomedical Engineering, Radiology, and Materials Science and Engineering, Case Western Reserve University
Plant viral nanoparticles (VNPs) are promising platforms for applications in biomedicine. Here, we describe the procedures for plant VNP propagation, purification, characterization, and bioconjugation. Finally, we show the application of VNPs for tumor homing and imaging using a mouse xenograft model and fluorescence imaging.
Published November 16, 2012. Keywords: Cancer Biology, Bioengineering, Biomedical Engineering, Molecular Biology, Virology, Oncology, Viral nanoparticles, bioconjugate chemistry, tumor xenograft mouse model, fluorescence imaging
1Institute for Biological and Medical Imaging (IBMI), Helmholtz Zentrum München, 2Faculty of Medicine, Technische Universität München
We provide herein a detailed description of the experimental protocol for imaging with a newly developed hand-held optoacoustic (photoacoustic) system for three-dimensional functional and molecular imaging in real time. The demonstrated powerful performance and versatility may define new application areas of the optoacoustic technology in preclinical research and clinical practice.
Published November 4, 2014. Keywords: Physiology, Optoacoustic tomography, photoacoustic imaging, hand-held probe, volumetric imaging, real-time tomography, five dimensional imaging, clinical imaging, functional imaging, molecular imaging, preclinical research
1ILM-FENNEC UMR 5306, CNRS - Université Lyon 1, 2ILM-PUBLI UMR 5306, CNRS - Université Lyon 1, 3ILM-SOPRANO UMR 5306, CNRS - Université Lyon 1
Laser-induced breakdown spectroscopy performed on thin organ and tumor tissue successfully detected natural elements and artificially injected gadolinium (Gd), issued from Gd-based nanoparticles. Images of chemical elements reached a resolution of 100 μm and quantitative sub-mM sensitivity. The compatibility of the setup with standard optical microscopy emphasizes its potential to provide multiple images of a same biological tissue.
Published June 18, 2014. Keywords: Physics, Microtechnology, Nanotechnology, Tissues, Diagnosis, Inorganic Chemistry, Organic Chemistry, Physical Chemistry, Plasma Physics, laser-induced breakdown spectroscopy, nanoparticles, elemental mapping, chemical images of organ tissue, quantification, biomedical measurement, laser-induced plasma, spectrochemical analysis, tissue mapping
1Division of Developmental Biology, Cincinnati Children's Hospital Medical Center, 2Division of Hematology/Oncology, Cincinnati Children's Hospital Medical Center
A method is described to photoactivate single cells containing a caged fluorescent protein using two-photon absorption from a Ti:Sapphire femtosecond laser oscillator. To fate map the photoactivated cell, immunohistochemistry is used. This technique can be applied to any cell type.
Published October 5, 2011. Keywords: Developmental Biology, Zebrafish, Two-photon, photoactivation, immunohistochemistry, caged fluorescent protein, fate mapping
1Molecular Biology and Biochemistry, University of California, Irvine, 2Physiology and Biophysics, University of California, Irvine, 3Neurobiology and Behavior, University of California, Irvine, 4University of California San Francisco Diabetes Center, University of California, San Francisco, 5Pathology, University of Utah
A new ex vivo preparation for imaging the mouse spinal cord. This protocol allows for two-photon imaging of live cellular interactions throughout the spinal cord.
Published February 22, 2015. Keywords: Neuroscience, spinal cord, two-photon microscopy, ex vivo, transplantation, cellular dynamics, axons, neural precursor cells, remyelination, neuroinflammation
1Department of Urology, Academic Medical Center, 2Department of Biomedical Engineering and Physics, Academic Medical Center, 3Department of Radiology, Academic Medical Center
Optical coherence tomography (OCT) is a high resolution imaging technique that allows analysis of tissue specific optical properties providing the means for tissue differentiation. We developed needle based OCT, providing real-time imaging combined with on-the-spot tumor differentiation. This publication describes a method for percutaneous, needle based OCT of renal masses.
Published March 30, 2015. Keywords: Medicine, Optical Coherence Tomography, OCT, Optical frequency domain imaging, OFDI, Optical biopsy, Needle based, Percutaneous, Renal mass, Kidney tumor, Kidney cancer.
1Division of Hematology-Oncology, Department of Internal Medicine, University of Cincinnati College of Medicine, 2Division of Human Genetics, University of Cincinnati College of Medicine, 3Department of Radiology, University of Cincinnati College of Medicine, 4Division of Rheumatology, Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, 5Solid Tumor Biology Program, James Comprehensive Cancer Center, The Ohio State University Medical Center, 6Department of Neurological Surgery, James Comprehensive Cancer Center, The Ohio State University Medical Center
We describe a multi-angle rotational optical imaging (MAROI) system for in vivo quantitation of a fluorescent marker delivered by saposin C (SapC)-dioleoylphosphatidylserine (DOPS) nanovesicles. Employing mouse models of cancer and arthritis, we demonstrate how the MAROI signal curve analysis can be used for the precise mapping and biological characterization of disease processes.
Published May 2, 2014. Keywords: Medicine, Saposin C (SapC), Dioleoylphosphatidylserine (DOPS), Brain tumor, Arthritis, Fluorophore, Fluorescence, Optical imaging, Multi-angle rotational optical imaging (MAROI)
1Center for Regenerative Medicine, Massachusetts General Hospital, Harvard Stem Cell Institute, 2Wellman Center for Photomedicine and Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School
Quantitative measurement of bone progenitor function in fracture healing requires high resolution serial imaging technology. Here, protocols are provided for using intravital microscopy and osteo-lineage tracking to sequentially image and quantify the migration, proliferation and differentiation of endogenous osteogenic stem/progenitor cells in the process of repairing bone fracture.
Published May 23, 2014. Keywords: Medicine, Osteogenic Stem Cells, In vivo Imaging, Lineage tracking, Bone regeneration, Fracture repair, Mx1.
1Bioengineering Department, Northeastern University, 2Department of Pharmaceutical Sciences, Northeastern University
Fluorescent nanoparticles produced in our lab are used for imaging ion concentrations and ion fluxes in biological systems such as cells during signaling and interstitial fluid during physiological homeostasis.
Published July 4, 2011. Keywords: Bioengineering, nanoparticles, nanosensors, polymer, fluorescence, imaging, intracellular, in vivo, sodium
JoVE Immunology and Infection
1Lurie Family Imaging Center, Dana-Farber Cancer Institute, Harvard Medical School, 2Division of Pediatric Hematology/Oncology/Stem Cell Transplantation, Columbia University Medical Center
We describe a non-invasive imaging method for distinguishing inflammatory stages. Systemic delivery of luminol reveals areas of acute inflammation dependent upon MPO activity in neutrophils. In contrast, injection of lucigenin allows for visualization of chronic inflammation dependent upon Phox activity in macrophages.
Published August 16, 2013. Keywords: Immunology, Infection, Medicine, Cellular Biology, Molecular Biology, Biomedical Engineering, Anatomy, Physiology, Cancer Biology, Stem Cell Biology, Inflammation, Phagocytes, Phagocyte, Superoxides, Molecular Imaging, chemiluminescence, in vivo imaging, superoxide, bioluminescence, chronic inflammation, acute inflammation, phagocytes, cells, imaging, animal model
1Department of Biomedical Engineering, University at Buffalo, The State University of New York, 2Department of Creative IT Engineering, Pohang University of Science and Technology (POSTECH), 3School of Electrical Engineering and Computer Science, Kyungpook National University
Photoacoustic cystography (PAC) has a great potential to map urinary bladders, a radiation sensitive internal organ in pediatric patients, without using any ionizing radiation or toxic contrast agent. Here we demonstrate the use of PAC for mapping urinary bladders with an injection of optical-opaque tracers in rats in vivo.
Published June 11, 2013. Keywords: Biomedical Engineering, Biophysics, Medicine, Bioengineering, Cancer Biology, Engineering (General), Electronics and Electrical Engineering, Lasers and Masers, Acoustics, Optics, Photoacoustic cystography, nonionizing imaging, contrast agent, urinary tract reflux, bladder, cystography, photoacoustic tomography, PAT, tomography, imaging, clinical techniques, animal model