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MGH - Massachusetts General Hospital

15 ARTICLES PUBLISHED IN JoVE

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Biology

Horizontal Slice Preparation of the Retina
Ryosuke Enoki 1, Tatjana C. Jakobs 2, Amane Koizumi 2
1Dpt of Physiology and Biophysics, Dalhousie University, 2Massachusetts General Hospital, Harvard Medical School

Traditionally the vertical slice and the whole-mount preparation of the retina have been used to study the function of retinal circuits. Here, we describe the novel slicing method to preserve the dendritic morphology of retinal neurons intact.

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Biology

Propagation of Human Embryonic Stem (ES) Cells
Laurence Daheron 1
1Center for Regenerative Medicine, MGH - Massachusetts General Hospital

Propagation of Human Embryonic Stem (ES) Cells

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Biology

Organotypic Culture of Adult Rabbit Retina
Ming H. Lye 1, Tatjana C. Jakobs 1, Richard H. Masland 1, Amane Koizumi 1
1Havard Medical School, MGH - Massachusetts General Hospital

This article demonstrates the dissection and incubation of rabbit retina and particle-mediated gene transfer of plasmids encoding GFP or a variety of subcellular markers into retinal ganglion cells.

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Biology

PDMS Device Fabrication and Surface Modification
Kenneth Kotz 1, Xuanhong Cheng 1, Mehmet Toner 1
1Havard Medical School, MGH - Massachusetts General Hospital

PDMS Device Fabrication and Surface Modification

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Biology

Cell Capture Using a Microfluidic Device
Kenneth Kotz 1, Xuanhong Cheng 1, Mehmet Toner 1
1Havard Medical School, MGH - Massachusetts General Hospital

Cell Capture Using a Microfluidic Device

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Biology

Development of New Therapeutic Applications Using Microfluidics
Mehmet Toner 1
1Havard Medical School, MGH - Massachusetts General Hospital

Development of New Therapeutic Applications Using Microfluidics

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Biology

Homing of Hematopoietic Cells to the Bone Marrow
Rushdia Z. Yusuf 1, David T. Scadden 1
1Center for Regenerative Medicine, MGH - Massachusetts General Hospital

This article describes a protocol used to study the homing of hematopoietic cells to their niches in the bone marrow.

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Biology

Born Normalization for Fluorescence Optical Projection Tomography for Whole Heart Imaging
Claudio Vinegoni 1,2, Daniel Razansky 3, Jose-Luiz Figueiredo 1,2, Lyuba Fexon 1,2, Misha Pivovarov 1,2, Matthias Nahrendorf 1,2, Vasilis Ntziachristos 3, Ralph Weissleder 1,2
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.

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Biology

Mesoscopic Fluorescence Tomography for In-vivo Imaging of Developing Drosophila
Claudio Vinegoni 1, Daniel Razansky 2, Chrysoula Pitsouli 3, Norbert Perrimon 3, Vasilis Ntziachristos 2, Ralph Weissleder 1
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.

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Neuroscience

Functional Mapping with Simultaneous MEG and EEG
Hesheng Liu 1, Naoaki Tanaka 1, Steven Stufflebeam 1, Seppo Ahlfors 1, Matti Hämäläinen 1
1Athinoula A. Martinos Center for Biomedical Imaging, MGH - Massachusetts General Hospital

We use magnetoencephalography (MEG) and electroencephalography (EEG) to map brain areas involved in the processing of simple sensory stimuli.

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Chemistry

Microfluidic On-chip Capture-cycloaddition Reaction to Reversibly Immobilize Small Molecules or Multi-component Structures for Biosensor Applications
Carlos Tassa 1, Monty Liong 1, Scott Hilderbrand 1, Jason E. Sandler 1, Thomas Reiner 1, Edmund J. Keliher 1, Ralph Weissleder 1, Stanley Y. Shaw 1
1Center for Systems Biology, Massachusetts General Hospital

We present a method for rapid, reversible immobilization of small molecules and functionalized nanoparticle assemblies for Surface Plasmon Resonance (SPR) studies, using sequential on-chip bioorthogonal cycloaddition chemistry and antibody-antigen capture.

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Medicine

Sequential In vivo Imaging of Osteogenic Stem/Progenitor Cells During Fracture Repair
Dongsu Park 1, Joel A. Spencer 2, Charles P. Lin 2, David T. Scadden 1
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.

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JoVE Core

Interictal High Frequency Oscillations Detected with Simultaneous Magnetoencephalography and Electroencephalography as Biomarker of Pediatric Epilepsy
Christos Papadelis 1, Eleonora Tamilia 1, Steven Stufflebeam 2, Patricia E. Grant 1, Joseph R. Madsen 3, Phillip L. Pearl 4, Naoaki Tanaka 2
1Fetal-Neonatal Neuroimaging and Developmental Science Center, Division of Newborn Medicine, Department of Medicine, Boston Children's Hospital, Harvard Medical School, 2Athinoula A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital, Harvard Medical School, 3Division of Epilepsy Surgery, Department of Neurosurgery, Boston Children's Hospital, Harvard Medical School, 4Division of Epilepsy and Clinical Neurophysiology, Department of Neurology, Boston Children's Hospital, Harvard Medical School

High Frequency Oscillations (HFOs) have emerged as presurgical biomarkers for the identification of the epileptogenic zone in pediatric patients with medically refractory epilepsy. A methodology for the noninvasive recording, detection, and localization of HFOs with simultaneous scalp electroencephalography (EEG) and magnetoencephalography (MEG) is presented.

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Bioengineering

Bulk Droplet Vitrification for Primary Hepatocyte Preservation
Reinier J. de Vries 1,2,3, Peony D. Banik 1,2, Sonal Nagpal 1,2, Lindong Weng 1,2, Sinan Ozer 1,2, Thomas M. van Gulik 3, Mehmet Toner 1,2, Shannon N. Tessier 1,2, Korkut Uygun 1,2
1Center for Engineering in Medicine, Harvard Medical School, Massachusetts General Hospital, 2Shriners Hospitals for Children, Boston, 3Department of Surgery, University of Amsterdam

This manuscript describes an ice-free cryopreservation method for large quantities of rat hepatocytes whereby primary cells are pre-incubated with cryoprotective agents at a low concentration and vitrified in large droplets.

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Bioengineering

Characterizing Single-Molecule Conformational Changes Under Shear Flow with Fluorescence Microscopy
Avani V. Pisapati 1, Yi Wang *2, Megan E. Blauch *3, Nathan J. Wittenberg 3, Xuanhong Cheng 1,2, X. Frank Zhang 1,4
1Department of Bioengineering, Lehigh University, 2Department of Materials Science and Engineering, Lehigh University, 3Department of Chemistry, Lehigh University, 4Department of Mechanical Engineering and Mechanics, Lehigh University

We present a protocol for immobilizing single macromolecules in microfluidic devices and quantifying changes in their conformations under shear flow. This protocol is useful for characterizing the biomechanical and functional properties of biomolecules such as proteins and DNA in a flow environment.

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