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Beth Israel Deaconess Medical Center

25 ARTICLES PUBLISHED IN JoVE

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Biology

Obtaining High Quality RNA from Single Cell Populations in Human Postmortem Brain Tissue
Charmaine Y. Pietersen 1, Maribel P. Lim 1, Tsung-Ung W. Woo 1,2,3
1Department of Structural and Molecular Neuroscience, McLean Hospital, 2Department of Psychiatry, Harvard Medical School, 3Department of Psychiatry, Beth Israel Deaconess Medical Center

We describe a process using laser-capture microdissection to isolate and extract RNA from a homogeneous cell population, pyramidal neurons, in layer III of the superior temporal gyrus in postmortem human brains. We subsequently linearly amplify (T7-based) mRNA, and hybridize the sample to the Affymetrix human X3P microarray.

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Neuroscience

A Novel Approach for Documenting Phosphenes Induced by Transcranial Magnetic Stimulation
Seth Elkin-Frankston 1, Peter J. Fried 1, Alvaro Pascual-Leone 2, R. J. Rushmore III 1, Antoni Valero-Cabré 1,3
1Department of Anatomy and Neurobiology, Boston University School of Medicine, 2Department of Neurology, Beth Israel Deaconess Med Center, 3Centre de Recherche de l'institut du Cerveau et la Moelle Epinière (CRICM), Centre National de la Recherche Scientifique (CNRS)

Phosphenes are transient percepts of light that can be induced by applying Transcranial Magnetic Stimulation (TMS) to visually sensitive regions of cortex. We demonstrate a standard protocol for determining the phosphene threshold value and introduce a novel method for quantifying and analyzing perceived phosphenes.

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Neuroscience

Combining Transcranial Magnetic Stimulation and fMRI to Examine the Default Mode Network
Mark A. Halko 1, Mark C. Eldaief 1, Jared C. Horvath 1, Alvaro Pascual-Leone 1
1Berenson-Allen Center for Noninvasive Brain Stimulation, Beth Israel Deaconess Medical Center

In this article, we examine the methodology and considerations relevant to the combination of TMS and fMRI to examine the effects of brain stimulation on the default network.

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Neuroscience

TMS: Using the Theta-Burst Protocol to Explore Mechanism of Plasticity in Individuals with Fragile X Syndrome and Autism
Lindsay M. Oberman 1, Jared C. Horvath 1, Alvaro Pascual-Leone 1
1Berenson-Allen Center for Noninvasive Brain Stimulation, Beth Israel Deaconess Medical Center

In this article, we examine the effects of Theta-Burst TMS stimulation on cortical plasticity in individuals suffering from Fragile X syndrome and individuals on the autistic spectrum.

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Neuroscience

State-Dependency Effects on TMS: A Look at Motive Phosphene Behavior
Umer Najib 1, Jared C. Horvath 1, Juha Silvanto 2, Alvaro Pascual-Leone 1
1Berenson-Allen Center for Noninvasive Brain Stimulation, Beth Israel Deaconess Medical Center, 2Brain Research Unit, Low Temperature Laboratory and Advanced magnetic Imaging Center, Aalto University School of Science and Technology

In this article, we examine the effects of visually relevant state dependency on TMS induced motive phosphenic presentations.

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Neuroscience

The NeuroStar TMS Device: Conducting the FDA Approved Protocol for Treatment of Depression
Jared C. Horvath 1, John Mathews 2, Mark A. Demitrack 2, Alvaro Pascual-Leone 1
1Berenson-Allen Center for Noninvasive Brain Stimulation, Beth Israel Deaconess Medical Center, 2Neuronetics, Inc.

In this article, we examine the methodology and considerations relevant to the FDA approved depression treatment protocol using the Neuronetics NeuroStar TMS device.

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Neuroscience

Generation of Neural Stem Cells from Discarded Human Fetal Cortical Tissue
Jie Lu 1, Laurent C. Delli-Bovi 2, Jonathan Hecht 3, Rebecca Folkerth 4, Volney L. Sheen 1
1Department of Neurology, Beth Israel Deaconess Medical Center, 2Department of Obstetrics and Gynecology, Brigham and Women's Hospital, 3Department of Pathology, Beth Israel Deaconess Medical Center, 4Department of Pathology, Division of Neuropathology, Brigham and Women's Hospital

A simple and reliable method on isolation and culture of neural stem cells from discarded human fetal cortical tissue is described. Cultures derived from known human neurological disorders can be used for characterization of pathological cellular and molecular processes, as well as provide a platform to assess pharmacological efficacy.

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Clinical Skills

Respiratory Exam I: Inspection and Palpation
Suneel Dhand 1
1Beth Israel Deaconess Medical Center

Respiratory Exam I: Inspection and Palpation

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Clinical Skills

Respiratory Exam II: Percussion and Auscultation
Suneel Dhand 1
1Beth Israel Deaconess Medical Center

Respiratory Exam II: Percussion and Auscultation

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Clinical Skills

Cardiac Exam I: Inspection and Palpation
Suneel Dhand 1
1Beth Israel Deaconess Medical Center

Cardiac Exam I: Inspection and Palpation

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Clinical Skills

Blood Pressure Measurement
Megan Fashijian 1
1Beth Israel Deaconess Medical Center

Blood Pressure Measurement

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Clinical Skills

Abdominal Exam I: Inspection and Auscultation
Alex Goldfarb 1
1Beth Israel Deaconess Medical Center

Abdominal Exam I: Inspection and Auscultation

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Clinical Skills

Abdominal Exam III: Palpation
Alex Goldfarb 1
1Beth Israel Deaconess Medical Center

Abdominal Exam III: Palpation

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Clinical Skills

Abdominal Exam II: Percussion
Alex Goldfarb 1
1Beth Israel Deaconess Medical Center

Abdominal Exam II: Percussion

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Clinical Skills

Measuring Vital Signs
Megan Fashjian 1
1Beth Israel Deaconess Medical Center

Measuring Vital Signs

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Clinical Skills

Cardiac Exam II: Auscultation
Suneel Dhand 1
1Beth Israel Deaconess Medical Center

Cardiac Exam II: Auscultation

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Clinical Skills

Cardiac Exam III: Abnormal Heart Sounds
Suneel Dhand 1
1Beth Israel Deaconess Medical Center

Cardiac Exam III: Abnormal Heart Sounds

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Medicine

A Multimodal Imaging- and Stimulation-based Method of Evaluating Connectivity-related Brain Excitability in Patients with Epilepsy
Mouhsin M. Shafi 1,2,3, Susan Whitfield-Gabrieli 4, Catherine J. Chu 1,5, Alvaro Pascual-Leone 1,2,3, Bernard S. Chang 1,2
1Department of Neurology, Harvard Medical School, 2Department of Neurology, Beth Israel Deaconess Medical Center, 3Berenson-Allen Center for Noninvasive Brain Stimulation, Beth Israel Deaconess Medical Center, 4Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, 5Department of Neurology, Massachusetts General Hospital

Resting-state functional-connectivity MRI has identified abnormalities in patients with a wide range of neuropsychiatric disorders, including epilepsy due to malformations of cortical development. Transcranial Magnetic Stimulation in combination with EEG can demonstrate that patients with epilepsy have cortical hyperexcitability in regions with abnormal connectivity.

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Immunology and Infection

High Throughput, Real-time, Dual-readout Testing of Intracellular Antimicrobial Activity and Eukaryotic Cell Cytotoxicity
Lucius Chiaraviglio 1, Yoon-Suk Kang 1, James E. Kirby 1
1Department of Pathology, Beth Israel Deaconess Medical Center

A high throughput, real-time assay was developed to simultaneously identify (1) eukaryotic cell-penetrant antimicrobials targeting an intracellular bacterial pathogen, and (2) assess eukaryotic cell cytotoxicity. A variation on the same technology was thereafter combined with digital dispensing technology to enable facile, high-resolution, dose-response, and two- and three-dimensional synergy studies.

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Immunology and Infection

Antimicrobial Synergy Testing by the Inkjet Printer-assisted Automated Checkerboard Array and the Manual Time-kill Method
Thea Brennan-Krohn 1,2,3, James E Kirby 1,3
1Department of Pathology, Beth Israel Deaconess Medical Center, 2Division of Infectious Diseases, Boston Children's Hospital, 3Harvard Medical School

Antimicrobial synergy testing is used to evaluate the effect of two or more antibiotics used in combination and is typically performed by one of two methods: the checkerboard array or the time-kill assay. Here, we present an automated, inkjet printer-assisted checkerboard array synergy technique and a classic time-kill synergy study.

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Bioengineering

High Throughput Traction Force Microscopy Using PDMS Reveals Dose-Dependent Effects of Transforming Growth Factor-β on the Epithelial-to-Mesenchymal Transition
Haruka Yoshie 1, Newsha Koushki 1, Clayton Molter 1, Peter M. Siegel 2,3, Ramaswamy Krishnan 4, Allen J. Ehrlicher 1,2
1Department of Bioengineering, McGill University, 2Goodman Cancer Research Centre, McGill University, 3Department of Medicine, McGill University, 4Department of Emergency Medicine, Beth Israel Deaconess Medical Center

We present a high throughput traction force assay fabricated with silicone rubber (PDMS). This novel assay is suitable for studying physical changes in cell contractility during various biological and biomedical processes and diseases. We demonstrate this method's utility by measuring a TGF-β dependent increase in contractility during the epithelial-to-mesenchymal transition.

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Biochemistry

Atomic Absorbance Spectroscopy to Measure Intracellular Zinc Pools in Mammalian Cells
Shellaina J.V. Gordon *1, Yao Xiao *2, Amanda L. Paskavitz 3, Napoleón Navarro-Tito 4, Juan G. Navea 2, Teresita Padilla-Benavides 1
1Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School, 2Department of Chemistry, Skidmore College, 3Candiac MR Center, Beth Israel Deaconess Medical Center, 4Facultad de Ciencias Químico Biológicas, Universidad Autónoma de Guerrero

Cultured primary or established cell lines are commonly used to address fundamental biological and mechanistic questions as an initial approach before using animal models. This protocol describes how to prepare whole cell extracts and subcellular fractions for studies of zinc (Zn) and other trace elements with atomic absorbance spectroscopy.

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Neuroscience

Derivation, Expansion, Cryopreservation and Characterization of Brain Microvascular Endothelial Cells from Human Induced Pluripotent Stem Cells
Sovannarath Pong 1,2,3, Paulo Lizano 1,2,3,4, Rakesh Karmacharya 1,3,4,5
1Center for Genomic Medicine, Massachusetts General Hospital, 2Department of Psychiatry, Beth Israel Deaconess Medical Center, 3Chemical Biology and Therapeutic Science Program, Broad Institute of MIT and Harvard, 4Department of Psychiatry, Harvard Medical School, 5Schizophrenia and Bipolar Disorder Program, McLean Hospital

This protocol details an adapted method to derive, expand, and cryopreserve brain microvascular endothelial cells obtained by differentiating human induced pluripotent stem cells, and to study blood brain barrier properties in an ex vivo model.

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Biology

Quantification of Cellular Densities and Antigenic Properties using Magnetic Levitation
Lauren Thompson 1, Brandy Pinckney 1, Shulin Lu 2, Mark Gregory 2, John Tigges 1, Ionita Ghiran 2
1Nano Flow Core Facility, Beth Israel Deaconess Medical Center, Harvard Medical School, 2Department of Medicine, Division of Allergy and Inflammation, Beth Israel Deaconess Medical Center

This paper describes a magnetic levitation-based method that can specifically detect the presence of antigens, either soluble or membrane-bound, by quantifying changes in the levitation height of capture beads with fixed densities.

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Neuroscience

Manual Segmentation of the Human Choroid Plexus Using Brain MRI
Deepthi Bannai *1,2, Yuan Cao *3,4, Matcheri Keshavan 1,2, Martin Reuter 5,6,7, Paulo Lizano 1,2,8
1Department of Psychiatry, Beth Israel Deaconess Medical Center, 2Division of Translational Neuroscience, Beth Israel Deaconess Medical Center, 3Huaxi MR Research Center (HMRRC), Department of Radiology, West China Hospital of Sichuan University, 4Department of Psychiatry and Psychotherapy, Jena University Hospital, 5AI in Medical Imaging, German Center for Neurodegenerative Diseases (DZNE), 6A. A. Martinos Center for Biomedical Imaging, Massachusetts General Hospital, 7Department of Radiology, Harvard Medical School, 8Department of Psychiatry, Harvard Medical School

Despite the crucial role of the choroid plexus in the brain, neuroimaging studies of this structure are scarce due to the lack of reliable automated segmentation tools. The present protocol aims to ensure gold-standard manual segmentation of the choroid plexus that can inform future neuroimaging studies.

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