Beth Israel Deaconess Medical Center View Institution's Website 13 articles published in JoVE Biology Quantification of Cellular Densities and Antigenic Properties using Magnetic Levitation Lauren Thompson1, Brandy Pinckney1, Shulin Lu2, Mark Gregory2, John Tigges1, Ionita Ghiran2 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. Neuroscience Derivation, Expansion, Cryopreservation and Characterization of Brain Microvascular Endothelial Cells from Human Induced Pluripotent Stem Cells Sovannarath Pong1,2,3, Paulo Lizano1,2,3,4, Rakesh Karmacharya1,3,4,5 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. Bioengineering High Throughput Traction Force Microscopy Using PDMS Reveals Dose-Dependent Effects of Transforming Growth Factor-β on the Epithelial-to-Mesenchymal Transition Haruka Yoshie1, Newsha Koushki1, Clayton Molter1, Peter M. Siegel2,3, Ramaswamy Krishnan4, Allen J. Ehrlicher1,2 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. Immunology and Infection Antimicrobial Synergy Testing by the Inkjet Printer-assisted Automated Checkerboard Array and the Manual Time-kill Method Thea Brennan-Krohn1,2,3, James E Kirby1,3 1Department of Pathology, Beth Israel Deaconess Medical Center, 2 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. Immunology and Infection High Throughput, Real-time, Dual-readout Testing of Intracellular Antimicrobial Activity and Eukaryotic Cell Cytotoxicity Lucius Chiaraviglio1, Yoon-Suk Kang1, James E. Kirby1 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. Medicine A Multimodal Imaging- and Stimulation-based Method of Evaluating Connectivity-related Brain Excitability in Patients with Epilepsy Mouhsin M. Shafi1,2,3, Susan Whitfield-Gabrieli4, Catherine J. Chu1,5, Alvaro Pascual-Leone1,2,3, Bernard S. Chang1,2 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. Neuroscience Generation of Neural Stem Cells from Discarded Human Fetal Cortical Tissue Jie Lu1, Laurent C. Delli-Bovi2, Jonathan Hecht3, Rebecca Folkerth4, Volney L. Sheen1 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. Neuroscience State-Dependency Effects on TMS: A Look at Motive Phosphene Behavior Umer Najib1, Jared C. Horvath1, Juha Silvanto2, Alvaro Pascual-Leone1 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. Neuroscience TMS: Using the Theta-Burst Protocol to Explore Mechanism of Plasticity in Individuals with Fragile X Syndrome and Autism Lindsay M. Oberman1, Jared C. Horvath1, Alvaro Pascual-Leone1 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. Neuroscience Combining Transcranial Magnetic Stimulation and fMRI to Examine the Default Mode Network Mark A. Halko1, Mark C. Eldaief1, Jared C. Horvath1, Alvaro Pascual-Leone1 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. Neuroscience The NeuroStar TMS Device: Conducting the FDA Approved Protocol for Treatment of Depression Jared C. Horvath1, John Mathews2, Mark A. Demitrack2, Alvaro Pascual-Leone1 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. Neuroscience A Novel Approach for Documenting Phosphenes Induced by Transcranial Magnetic Stimulation Seth Elkin-Frankston1, Peter J. Fried1, Alvaro Pascual-Leone2, R. J. Rushmore III1, Antoni Valero-Cabré1,3 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. Biology Obtaining High Quality RNA from Single Cell Populations in Human Postmortem Brain Tissue Charmaine Y. Pietersen1, Maribel P. Lim1, Tsung-Ung W. Woo1,2,3 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.