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Massachusetts General Hospital and Harvard Medical School

24 ARTICLES PUBLISHED IN JoVE

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Biology

Monitoring Acupuncture Effects on Human Brain by fMRI
Kathleen K. S. Hui 1, Vitaly Napadow 1, Jing Liu 1, Ming Li 1, Ovidiu Marina 1,2, Erika E. Nixon 1, Joshua D. Claunch 1, Lauren LaCount 1, Tara Sporko 1, Kenneth K. Kwong 1
1Department of Radiology, Massachusetts General Hospital and Harvard Medical School, 2William Beaumont Hospital

FMRI and physiological monitoring is used to study the effects of Acupuncture on the central and peripheral nervous systems. Acupuncture mobilizes a limbic-paralimbic-neocortical network, with great overlap with the default mode network, to modulate neurological activity, possibly related to its autonomic effect in the peripheral nervous system.

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Biology

Bioluminescence Imaging of Heme Oxygenase-1 Upregulation in the Gua Sha Procedure
Kenneth K. Kwong 1,2, Lenuta Kloetzer 1,2,3,4, Kelvin K. Wong 5,6, Jia-Qian Ren 1,2, Braden Kuo 1,2,3,4, Yan Jiang 7, Y. Iris Chen 1,2, Suk-Tak Chan 1,2,8, Geoffrey S. Young 9, Stephen T.C. Wong 5,6
1Department of Radiology, Massachusetts General Hospital, Harvard Medical School, 2Athinoula A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital, Harvard Medical School, 3Gastrointestinal Unit, Massachusetts General Hospital, Harvard Medical School, 4Department of Medicine, Massachusetts General Hospital, Harvard Medical School, 5Center for biotechnology and Informatics, The Methodist Hospital Research Institute, 6Department of Radiology, The Methodist Hospital, Weill Cornell Medical College, 7Bejing University of Chinese Medicine, 8Department of Health Technology and Informatics, The Hong Kong Polytechnic University, 9Department of Radiology, Brigham and Women's Hospital, Harvard Medical School

Gua Sha, traditional Chinese therapeutic skin scraping, causes subcutaneous microvascular blood extravasation. We report a protocol of bioluminescence imaging of HO-1-luciferase transgenic mice to demonstrate that Gua Sha upregulates heme oxygenase-1 (HO-1) in multiple organs.

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Neuroscience

Cerebral Blood Oxygenation Measurement Based on Oxygen-dependent Quenching of Phosphorescence
Sava Sakadžić 1, Emmanuel Roussakis 2, Mohammad A. Yaseen 1, Emiri T. Mandeville 3, Vivek J. Srinivasan 1, Ken Arai 3, Svetlana Ruvinskaya 1, Weicheng Wu 1, Anna Devor 1,4, Eng H. Lo 3, Sergei A. Vinogradov 2, David A. Boas 1
1Optics Division, Athinoula A. Martinos Center for Biomedical Imaging, Department of Radiology, Massachusetts General Hospital and Harvard Medical School, 2Department of Biochemistry and Biophysics, University of Pennsylvania, 3Neuroprotection Research Laboratory, Departments of Radiology and Neurology, Massachusetts General Hospital and Harvard Medical School, 4Departments of Neurosciences and Radiology, University of California

We present an experimental procedure for measuring the partial pressure of oxygen (pO2) in cerebral vasculature based on oxygen-dependent quenching of phosphorescence. Animal preparation and imaging procedures were outlined for both large field of view CCD-based imaging of pO2 in rats and 2-photon excitation based imaging of pO2 in mice.

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Immunology and Infection

Peptide:MHC Tetramer-based Enrichment of Epitope-specific T cells
Francois P. Legoux 1, James J. Moon 1
1Center for Immunology and Inflammatory Diseases, and Pulmonary and Critical Care Unit, Massachusetts General Hospital and Harvard Medical School

This protocol describes the use of peptide:MHC tetramers and magnetic microbeads to isolate low frequency populations of epitope-specific T cells and analyze them by flow cytometry. This method enables the direct study of endogenous T cell populations of interest from in vivo experimental systems.

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Education

Biochemical and High Throughput Microscopic Assessment of Fat Mass in Caenorhabditis Elegans
Elizabeth C. Pino 1, Christopher M. Webster 1, Christopher E. Carr 2, Alexander A. Soukas 1
1Center for Human Genetic Research and Department of Medicine, Massachusetts General Hospital and Harvard Medical School, 2Department of Earth, Atmospheric, and Planetary Sciences, Massachusetts Institute of Technology

We present robust biochemical and microscopic methods for studying Caenorhabditis elegans lipid stores. A rapid, simple, fixing-staining procedure for fluorescent lipid droplet imaging leverages the spectral properties of the lipophilic dye Nile red. We then present biochemical measurement of triglycerides and phospholipids using solid phase extraction and gas chromatography-mass spectrometry.

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Bioengineering

Generation of Aligned Functional Myocardial Tissue Through Microcontact Printing
Ayhan Atmanli 1, Ibrahim J. Domian 1,2
1Cardiovascular Research Center, Massachusetts General Hospital and Harvard Medical School, 2Harvard Stem Cell Institute

The generation of aligned myocardial tissue is a key requirement for adapting the recent advances in stem cell biology to clinically useful purposes. Herein we describe a microcontact printing approach for the precise control of cell shape and function. Using highly purified populations of embryonic stem cell derived cardiac progenitors, we then generate anisotropic functional myocardial tissue.

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Biology

Isolation and Culture of Adult Mouse Cardiomyocytes for Cell Signaling and in vitro Cardiac Hypertrophy
Daxiang Li 1, Jian Wu 1, Yan Bai 1, Xiaochen Zhao 2, Lijun Liu 1
1Department of Biochemistry and Cancer Biology, University of Toledo College of Medicine and Life Sciences, 2Department of Physiology and Pharmacology, University of Toledo College of Medicine and Life Sciences

We describe a reliable method for isolation of adult mouse cardiomyocytes. This protocol yields a consistent result for the culture of functional adult cardiomyocytes from a variety of genetically modified mice.

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JoVE Journal

Fabrication and Testing of Microfluidic Optomechanical Oscillators
Kewen Han 1, Kyu Hyun Kim 2, Junhwan Kim 1, Wonsuk Lee 2,3, Jing Liu 3, Xudong Fan 3, Tal Carmon 2, Gaurav Bahl 1
1Mechanical Science and Engineering, University of Illinois at Urbana-Champaign, 2Electrical Engineering and Computer Science, University of Michigan, 3Biomedical Engineering, University of Michigan

Parametric optomechanical excitations have recently been experimentally demonstrated in microfluidic optomechanical resonators by means of optical radiation pressure and stimulated Brillouin scattering. This paper describes the fabrication of these microfluidic resonators along with methodologies for generating and verifying optomechanical oscillations.

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Bioengineering

Cloning and Large-Scale Production of High-Capacity Adenoviral Vectors Based on the Human Adenovirus Type 5
Eric Ehrke-Schulz 1, Wenli Zhang 1, Maren Schiwon 1, Thorsten Bergmann 1, Manish Solanki 1, Jing Liu 1, Philip Boehme 1, Theo Leitner 1, Anja Ehrhardt 1
1Institute of Virology and Microbiology, Center for Biomedical Education and Research (ZBAF), Department of Human Medicine, Faculty of Health, University Witten/Herdecke

A protocol for generation of high-capacity adenoviral vectors lacking all viral coding sequences is presented. Cloning of transgenes contained in the vector genome is based on homing endonucleases. Virus amplification in producer cells grown as adherent cells and in suspension relies on a helper virus providing viral genes in trans.

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Immunology and Infection

SILAC Based Proteomic Characterization of Exosomes from HIV-1 Infected Cells
Collins Cheruiyot 1, Zemplen Pataki 1, Robert Williams 1, Bharat Ramratnam 2,3, Ming Li 3
1Brown University, 2COBRE Center for Cancer Research, Lifespan Laboratories, Rhode Island and Miriam Hospitals, 3Division of Infectious Diseases, Department of Medicine, Warren Alpert Medical School, Brown University

Here, we describe a quantitative proteomics method using the technique of stable isotope labeling by amino acids in cell culture (SILAC) to analyze the effects of HIV-1 infection on host exosomal proteomes. This protocol can be easily adapted to cells under different stress or infection conditions.

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Biology

Drosophila Preparation and Longitudinal Imaging of Heart Function In Vivo Using Optical Coherence Microscopy (OCM)
Jing Men 1,2, Jason Jerwick 2,3, Penghe Wu 1,2, Mingming Chen 3,4, Aneesh Alex 2,3, Yutao Ma 4, Rudolph E. Tanzi 5, Airong Li 5, Chao Zhou 1,2,3
1Bioengineering Program, Lehigh University, 2Center for Photonics and Nanoelectronics, Lehigh University, 3Department of Electrical and Computer Engineering, Lehigh University, 4State Key Laboratory of Software Engineering, Wuhan University, 5Genetics and Aging Research Unit, Department of Neurology, Massachusetts General Hospital and Harvard Medical School

Here, the experimental protocols are described for preparing Drosophila at different developmental stages and performing longitudinal optical imaging of Drosophila heartbeats using a custom optical coherence microscopy (OCM) system. The cardiac morphological and dynamical changes can be quantitatively characterized by analyzing the heart structural and functional parameters from OCM images.

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Genetics

Embryo Microinjection and Transplantation Technique for Nasonia vitripennis Genome Manipulation
Ming Li 1,2, Michelle Bui 1,2, Omar S. Akbari 1,2
1Department of Entomology and Riverside Center of Disease Vector Research, Institute for Integrative Genome Biology, University of California, Riverside, 2Section of Cell and Developmental Biology, Division of Biological Sciences, University of California, San Diego

Microinjection of Nasonia vitripennis embryos is an essential method for generating heritable genome modifications. Described here is a detailed procedure for microinjection and transplantation of Nasonia vitripennis embryos, which will greatly facilitate future genome manipulation in this organism.

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Neuroscience

Quantitative Analysis of Neuronal Dendritic Arborization Complexity in Drosophila
Shanshan Wang 1,2, Rudolph E. Tanzi 1, Airong Li 1
1Genetics and Aging Research Unit, Department of Neurology, Massachusetts General Hospital, Mass General Institute of Neurodegenerative Disease, 2Department of Geriatric Neurology, Nanlou Clinical Division, PLA General Hospital

This protocol focuses on quantitative analysis of neuronal dendritic arborization complexity (NDAC) in Drosophila, which can be used for studies of dendritic morphogenesis.

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Neuroscience

Neurovascular Network Explorer 2.0: A Simple Tool for Exploring and Sharing a Database of Optogenetically-evoked Vasomotion in Mouse Cortex In Vivo
Hana Uhlirova 1,2, Peifang Tian 3,4, Kıvılcım Kılıç 3,5, Martin Thunemann 1, Vishnu B. Sridhar 6, Radim Chmelik 2,7, Hauke Bartsch 1, Anders M. Dale 1,3, Anna Devor 1,3,8, Payam A. Saisan 3
1Department of Radiology, University of California, San Diego, 2Central European Institute of Technology, Brno University of Technology, 3Department of Neurosciences, University of California, San Diego, 4Department of Physics, John Carroll University, 5Department of Biomedical Engineering, Boston University, 6Bioengineering Undergraduate Program, University of California, San Diego, 7Institute of Physical Engineering, Faculty of Mechanical Engineering, Brno University of Technology, 8Martinos Center for Biomedical Imaging, Massachusetts General Hospital, Harvard Medical School

A graphical user interface for exploring and sharing a database of optogenetically-induced vascular responses in mouse somatosensory cortex in vivo measured by 2-photon microscopy is presented. It allows browsing the data, criteria-based selection, averaging, localization of measurements within a 3D volume of vasculature and exporting the data.

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Medicine

Endotoxin Activity Assay for the Detection of Whole Blood Endotoxemia in Critically Ill Patients
Riccardo Pinciroli 1,2, Simone Checchi 2, Maurizio Bottiroli 1, Gianpaola Monti 1, Giampaolo Casella 1, Roberto Fumagalli 1,2
1Department of Anesthesia and Critical Care, Niguarda Hospital, 2University of Milan-Bicocca Medical School

We hereby present a protocol to measure at the bedside the endotoxin activity of human whole blood samples. The Endotoxin Activity assay is a simple test to perform and may be a useful biomarker in critically ill patients with sepsis.

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Medicine

Protocol and Guidelines for Point-of-Care Lung Ultrasound in Diagnosing Neonatal Pulmonary Diseases Based on International Expert Consensus
Jing Liu 1,2, Roberto Copetti 3, Erich Sorantin 4, Jovan Lovrenski 5, Javier Rodriguez-Fanjul 6, Dalibor Kurepa 7, Xing Feng 8, Luigi Cattaross 9, Huayan Zhang 10,11, Misun Hwang 12, Tsu F. Yeh 13,14, Yisrael Lipener 7, Abhay Lodha 15, Jia-Qin Wang 16, Hai-Ying Cao 2,17, Cai-Bao Hu 2,18, Guo-Rong Lyu 19, Xin-Ru Qiu 1,2, Li-Qun Jia 20, Xiao-Man Wang 20, Xiao-Ling Ren 1,2, Jiu-Ye Guo 1,2, Yue-Qiao Gao 1,2, Jian-Jun Li 1,2, Ying Liu 1,2, Wei Fu 1,2, Yan Wang 21, Zu-Lin Lu 1,2, Hua-Wei Wang 8, Li-Li Shang 22
1Department of Neonatology and NICU, Beijing Chaoyang District Maternal and Child Healthcare Hospital, 2The Neonatal Lung Ultrasound Training Base, Chinese College of Critical Ultrasound, 3Emergency Department, Cattinara University Hospital, 4Division of Pediatric Radiology, Department of Radiology, Medical University Graz, 5Faculty of Medicine, University of Novi Sad, Radiology Department, Institute for Children and Adolescents Health Care of Vojvodina, 6Pediatric Intensive Care Unit, Pediatric Service Hospital Joan XXIII Tarragona, University Rovira i Virgil, 7Division of Neonatal-Perinatal Medicine, Cohen Children's Medical Center, 8Department of Neonatology, Children's Hospital of Soochow University, 9Department of Neonatology, Udine University Hospital, 10Center for Newborn Care, Guangzhou Women and Children's Medical Center, 11Division of Neonatology, Children's Hospital of Philadelphia, 12Department of Radiology, Children's Hospital of Philadelphia, 13Division of Neonatology and NICU, Cook County Children's Hospital, University of Illinois, 14Division of Neonatology, Department of Pediatrics, Taipei Medical University, 15Department of Pediatrics and Community Health Sciences, University of Calgary, 16Department of Pediatrics, The Third Affiliated Hospital of Xinxiang Medical University, 17Department of Ultrasound, GE Healthcare, 18Intensive Care Unit, Zhejiang Hospital, 19Collaborative Innovation Center for Maternal and Infant Health Service Application Technology, Quanzhou Medical College, 20Department of Ultrasound, Beijing Children's Hospital Affiliated with Capital Medical University, 21Department of Neonatology and NICU, Tai'an City Central Hospital of Shandong Province, 22Department of Intensive Care Unit, The Second Affiliated Hospital of Heilongjiang University of Chinese Medicine

Lung ultrasound is a noninvasive and valuable tool for bedside evaluation of neonatal lung diseases. However, a relative lack of reference standards, protocols and guidelines may limit its application. Here, we aim to develop a standardized neonatal lung ultrasound diagnostic protocol to be used in clinical decision-making.

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Neuroscience

Evaluation of Hemisphere Lateralization with Bilateral Local Field Potential Recording in Secondary Motor Cortex of Mice
Yunan Chen 1,2, Ming Li 3, Ying Zheng 3, Li Yang 1
1School of Life Sciences, Guangzhou University, 2Institute for Brain Research and Rehabilitation, South China Normal University, 3School of Life Sciences, South China Normal University

We present in vivo electrophysiological recording of the local field potential (LFP) in bilateral secondary motor cortex (M2) of mice, which can be applied to evaluate hemisphere lateralization. The study revealed altered levels of synchronization between the left and right M2 in APP/PS1 mice compared to WT controls.

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JoVE Core

International Expert Consensus and Recommendations for Neonatal Pneumothorax Ultrasound Diagnosis and Ultrasound-guided Thoracentesis Procedure
Jing Liu 1,2, Dalibor Kurepa 3, Francesco Feletti 4,5, Almudena Alonso-Ojembarrena 6, Jovan Lovrenski 7, Roberto Copetti 8, Erich Sorantin 9, Javier Rodriguez-Fanjul 10, Karishma Katti 3, Andrea Aliverti 4, Huayan Zhang 11,12, Misun Hwang 13, Tsu F. Yeh 14, Cai-Bao Hu 15, Xing Feng 16, Ru-Xin Qiu 1,2, Jing-Han Chi 17, Li-Li Shang 18, Guo-Rong Lyu 19, Shao-Zheng He 20, Yan-Fen Chai 21, Zhan-Jun Qiu 22, Hai-Ying Cao 2,23, Yue-Qiao Gao 1,2, Xiao-Ling Ren 1,2, Guo Guo 1,24, Li Zhang 1,2, Ying Liu 1,2, Wei Fu 1,2, Zu-Lin Lu 1,2, Hong-Lei Li 1,2
1Department of Neonatology and NICU, Beijing Chaoyang District Maternal and Child Healthcare Hospital, 2The National Neonatal Lung Ultrasound Training Base, 3Division of Neonatal-Perinatal Medicine, Cohen Children's Medical Center, 4Department of Electronics, Information and Bioengineering, Politecnico di Milano, 5Dipartimento di Diagnostica per Immagini, Ausl della Romagna, S. Maria delle Croci Hospital, 6Neonatal Intensive Care Unit, Puerta del Mar University Hospital, 7Faculty of Medicine, University of Novi Sad, Serbia, Institute for Children and Adolescents Health Care of Vojvodina, 8Emergency Department, University Hospital of Cattinara, 9Division of Pediatric Radiology, Department of Radiology, Medical University Graz, 10Pediatric Intensive Care Unit, Pediatric Service Hospital Joan XXIII Tarragona, University Rovira i Virgil, 11Center for Newborn Care, Guangzhou Women and Children's Medical Center, 12Division of Neonatology, Children's Hospital of Philadelphia, 13Section of Neonatal Imaging, Department of Radiology, Children's Hospital of Philadelphia, 14Maternal Child Health Research institute, Taipei Medical University and China Medical University, 15Intensive Care Unit, Zhejiang Hospital, 16Department of Neonatology, Children's Hospital of Soochow University, 17Department of Neonatology and NICU, Bayi Children's Hospital Affiliated to the Seventh Medical Center of Chinese PLA General Hospital, 18Intensive Care Unit, Second Affiliated Hospital of Heilongjiang University of Chinese Medicine, 19Collaborative Innovation Center for Maternal and Infant Health Service Application Technology, Quanzhou Medical College, 20Department of Ultrasound, Second Affiliated Hospital of Fujian Medical University, 21Department of Emergency Medicine, Tianjin Medical University General Hospital, 22Department of Emergency and Critical Care Medicine, Affiliated Hospital of Traditional Chinese Medicine, 23Department of Ultrasound, GE Healthcare, 24The Neonatal Intensive Care Unit, Fifth Medical Center of Chinese PLA General Hospital

Pneumothorax is a common emergency and critical disease in newborn infants that needs rapid, clear diagnosis and timely treatment. Diagnosis and treatment based on chest X-rays are associated with delayed management and radiation damage. Lung ultrasound (US) provides useful guidance for rapid, accurate diagnosis and the precise thoracentesis of pneumothorax.

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Genetics

Embryo Microinjection Techniques for Efficient Site-Specific Mutagenesis in Culex quinquefasciatus
Michelle Bui 1, Ming Li 1, Robyn R Raban 1, Nannan Liu 2, Omar S Akbari 1,3
1Section of Cell and Developmental Biology, University of California, San Diego, 2Department of Entomology and Plant Patholog, Auburn University, 3Tata Institute for Genetics and Society, University of California, San Diego

This protocol describes microinjection procedures for Culex quinquefasciatus embryos that are optimized to work with CRISPR/Cas9 gene editing tools. This technique can efficiently generate site-specific, heritable, germline mutations that can be used for building genetic technologies in this understudied disease vector.

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Medicine

A Novel Inhalation Mask System to Deliver High Concentrations of Nitric Oxide Gas in Spontaneously Breathing Subjects
Riccardo Pinciroli 1, Lisa Traeger 1, Anna Fischbach 1, Stefano Gianni 1, Caio Cesar Araujo Morais 1, Bijan Safaee Fakhr 1, Raffaele Di Fenza 1, Dianna Robinson 3, Ryan Carroll 2, Warren M. Zapol 1, Lorenzo Berra 1
1Department of Anesthesia, Critical Care and Pain Medicine, Massachusetts General Hospital and Harvard Medical School, 2Division of Pediatric Critical Care Medicine, Department of Pediatrics, Massachusetts General Hospital and Harvard Medical School, 3Respiratory Care, Massachusetts General Hospital

This simple and highly adaptable system device for the inhalation of high-concentration nitric oxide (NO) gas does not require mechanical ventilators, positive pressure, or high gas flows. Standard medical consumables and a snug-fitting mask are used to safely deliver NO gas to spontaneously breathing subjects.

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Immunology and Infection

Contact-Free Co-Culture Model for the Study of Innate Immune Cell Activation During Respiratory Virus Infection
Zhe Zhang Ryan Lew *1, Jing Liu *1, Hsiao Hui Ong 1, Vivian Jiayi Tan 2, Annika Luukkainen 3, Yew Kwang Ong 1,4, Mark Thong 1,4, Kia Joo Puan 5, Vincent Tak Kwong Chow 2,6, Kai Sen Tan 1,2,6, De Yun Wang 1,6
1Department of Otolaryngology, Yong Loo Lin School of Medicine, National University of Singapore, 2Department of Microbiology and Immunology, Yong Loo Lin School of Medicine, National University of Singapore, 3Haartman Institute, Medicum, University of Helsinki, 4Department of Otolaryngology, Head and Neck Surgery, National University Hospital, National University Health System, 5Agency for Science, Technology and Research (A*STAR), Singapore Immunology Network (SIgN), 6NUHS Infectious Diseases Translational Research Program, Yong Loo Lin School of Medicine, National University of Singapore

This protocol details an investigation of the early interactions between virally infected nasal epithelial cells and innate cell activation. Individual subsets of immune cells can be distinguished based on their activation in response to viral infections. They can then be further investigated to determine their effects on early antiviral responses.

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Immunology and Infection

Growth, Purification, and Titration of Oncolytic Herpes Simplex Virus
Hong-My Nguyen *1, Naresh Sah *1, Melissa R. M. Humphrey 2, Samuel D. Rabkin 2, Dipongkor Saha 1
1Department of Immunotherapeutics and Biotechnology, Jerry H. Hodge School of Pharmacy, Texas Tech University Health Sciences Center, 2Molecular Neurosurgery Laboratory and the Brain Tumor Research Center, Department of Neurosurgery, Massachusetts General Hospital and Harvard Medical School

In this manuscript, we describe a simple method of growth, purification, and titration of the oncolytic herpes simplex virus for preclinical use.

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Medicine

Chemiluminescence-based Assays for Detection of Nitric Oxide and its Derivatives from Autoxidation and Nitrosated Compounds
Raffaele Di Fenza 1,3, Binglan Yu 1,3, Ryan W. Carroll 2,3, Lorenzo Berra 1,3
1Department of Anesthesia, Critical Care and Pain Medicine, Massachusetts General Hospital, 2Department of Pediatrics, Massachusetts General Hospital, 3Harvard Medical School

Here, we present protocols for detecting nitric oxide and its biologically relevant derivatives using chemiluminescence-based assays with high sensitivity.

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Biology

Utilizing the Precision-Cut Lung Slice to Study the Contractile Regulation of Airway and Intrapulmonary Arterial Smooth Muscle
Yan Bai 1, Xingbin Ai 1
1Division of Newborn Medicine, Department of Pediatrics, Massachusetts General Hospital and Harvard Medical School

The present protocol describes preparing and utilizing mouse precision-cut lung slices to assess the airway and intrapulmonary arterial smooth muscle contractility in a nearly in vivo milieu.

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