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Brigham and Women's Hospital

24 ARTICLES PUBLISHED IN JoVE

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Biology

Fabrication of a Microfluidic Device for the Compartmentalization of Neuron Soma and Axons
Joseph Harris 1, Hyuna Lee 1, Behrad Vahidi 1, Christina Tu 2, David Cribbs 3, Noo Li Jeon 1, Carl Cotman 3
1Department of Biomedical Engineering, University of California, Irvine (UCI), 2Stem Cell Research Center, University of California, Irvine (UCI), 3Institute for Brain Aging and Dementia, University of California, Irvine (UCI)

In this video we demonstrate the technique of soft lithography with polydimethyl siloxane (PDMS) which we use to farbricate a microfluidic device for culturing neurons.

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Biology

A Microfluidic Device with Groove Patterns for Studying Cellular Behavior
Bong Geun Chung 1, Amir Manbachi 1, Ali Khademhosseini 1
1Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology; Center for Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital

We describe a protocol for the fabrication of microfluidic devices that can enable cell capture and culture. In this approach patterned microstructures such as grooves within microfluidic channels are used to create low shear stress regions within which cell can dock.

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Biology

A Gradient-generating Microfluidic Device for Cell Biology
Bong Geun Chung 1, Amir Manbachi 1, Wajeeh Saadi 1, Francis Lin 1, Noo Li Jeon 1, Ali Khademhosseini 1
1Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology; Center for Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital

We describe a protocol for the microfabrication of the gradient-generating microfluidic device that can generate spatial and temporal gradients in well-defined microenvironment. In this approach, the gradient-generating microfluidic device can be used to study directed cell migration, embryogenesis, wound healing, and cancer metastasis.

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Biology

Experimental Approaches to Tissue Engineering
Ali Khademhosseini 1
1Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology; Center for Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital

Experimental Approaches to Tissue Engineering

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Biology

Preparing E18 Cortical Rat Neurons for Compartmentalization in a Microfluidic Device
Joseph Harris 1, Hyuna Lee 1, Christina Tu Tu 2, David Cribbs 3, Carl Cotman 3, Noo Li Jeon 1
1Department of Biomedical Engineering, University of California, Irvine (UCI), 2Stem Cell Research Center, University of California, Irvine (UCI), 3Institute for Brain Aging and Dementia, University of California, Irvine (UCI)

In this video we demonstrate the preparation of E18 Cortical Rat Neurons.

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Biology

Using Micro-Electro-Mechanical Systems (MEMS) to Develop Diagnostic Tools
Utkan Demirci 1
1Bio-Acoustic-MEMS Laboratory in Medicine (BAMM), HST-Center for Bioengineering, Brigham and Women's Hospital

Using Micro-Electro-Mechanical Systems (MEMS) to Develop Diagnostic Tools

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Biology

CD4+ T-Lymphocyte Capture Using a Disposable Microfluidic Chip for HIV
Sang Jun Moon 1, Richard Lin 2, Utkan Demirci 1
1Bio-Acoustic-MEMS Laboratory in Medicine (BAMM), HST-Center for Bioengineering, Brigham and Women's Hospital, 2Massachusetts Institute of Technology

CD4+ T-Lymphocyte Capture Using a Disposable Microfluidic Chip for HIV

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Biology

Title Cell Encapsulation by Droplets
Sangjun Moon 1,2, Pei-Ann Lin 1,2, Hasan Onur Keles 1,2, Seung-Schick Yoo 3, Utkan Demirci 1,2,4
1Bio-Acoustic-MEMS Laboratory in Medicine (BAMM), HST-Center for Bioengineering, Brigham and Women's, Harvard Medical School, 2Bio-Acoustic-MEMS Laboratory in Medicine (BAMM), HST-Center for Bioengineering, Brigham and Women's Hospital, 3Brigham and Women's Hospital, Harvard Medical School, 4Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology; Center for Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital

Title Cell Encapsulation by Droplets

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Biology

Non-plasma Bonding of PDMS for Inexpensive Fabrication of Microfluidic Devices
Joseph Harris 1, Hyuna Lee 1, Behrad Vahidi 1, Cristina Tu 2, David Cribbs 3, Carl Cotman 3, Noo Li Jeon 1
1Department of Biomedical Engineering, University of California, Irvine (UCI), 2Stem Cell Research Center, University of California, Irvine (UCI), 3Institute for Brain Aging and Dementia, University of California, Irvine (UCI)

In this video we demonstrate how to use the neuron microfluidic device without plasma bonding.

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Biology

BioMEMS: Forging New Collaborations Between Biologists and Engineers
Noo Li Jeon 1
1Department of Biomedical Engineering, University of California, Irvine (UCI)

BioMEMS: Forging New Collaborations Between Biologists and Engineers

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Biology

Analysis of Physiologic E-Selectin-Mediated Leukocyte Rolling on Microvascular Endothelium
Georg Wiese 1, Steven R. Barthel 2, Charles J. Dimitroff 2
1Department of Dermatology, Brigham and Women's Hospital, 2Department of Dermatology, Brigham and Women's Hospital and Harvard Medical School

This report provides a visual depiction of parallel-plate flow chamber analysis for studying leukocyte endothelial interactions under physiologic shear stress. This method is particularly useful for investigating the role of endothelial (E)-selectin and leukocyte E-selectin ligands that trigger leukocyte rolling on endothelial cell surfaces.

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Biology

Mammary Epithelial Transplant Procedure
Karen A. Dunphy 1,2, Luwei Tao 3, D. Joseph Jerry 1,2
1Department of Veterinary and Animal Sciences, University of Massachussetts, 2Pioneer Valley Life Sciences institute, 3Molecular and Cellular Biology, University of Massachussetts

This article demonstrates the procedure developed by DeOme KB et al. (1959) and the sparing procedure developed by Brill B et al. (2008) for clearing the 4th inguinal mammary fat pad of a pubescent mouse in preparation for transplantation of mammary fragments, mammary epithelial cells, or mammary tumor cells.

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Medicine

Isolation of Mouse Respiratory Epithelial Cells and Exposure to Experimental Cigarette Smoke at Air Liquid Interface
Hilaire C. Lam 1,2, Augustine M.K. Choi 1, Stefan W. Ryter 1
1Department of Medicine, Pulmonary and Critical Care Medicine, Brigham and Women’s Hospital, Harvard Medical School, 2Cellular and Molecular Pathology, School of Medicine, University of Pittsburgh

Pulmonary epithelial cells can be isolated from the respiratory tract of mice and cultured at air-liquid interface as a model of differentiated respiratory epithelium. A protocol is described for isolating, culturing and exposing these cells to mainstream cigarette smoke, in order to study molecular responses to this environmental toxin.

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Bioengineering

Studying Cell Rolling Trajectories on Asymmetric Receptor Patterns
Chia-Hua Lee 1, Suman Bose 2, Krystyn J. Van Vliet 1, Jeffrey M. Karp 3, Rohit Karnik 2
1Department of Materials Science and Engineering, MIT - Massachusetts Institute of Technology, 2Department of Mechanical Engineering, MIT - Massachusetts Institute of Technology, 3HST Center for Biomedical Engineering and Harvard Stem Cell Institute, Brigham and Women's Hospital and Harvard Medical School

We describe a protocol to observe and analyze cell rolling trajectories on asymmetric receptor-patterned substrates. The resulting data are useful for engineering of receptor-patterned substrates for label-free cell separation and analysis.

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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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Biology

Ex Vivo Culture of Primary Human Fallopian Tube Epithelial Cells
Susan Fotheringham 1,2, Keren Levanon 1,2,3, Ronny Drapkin 1,2,4
1Department of Medical Oncology, Dana-Farber Cancer Institute, 2Harvard Medical School, Boston, MA, 3Sheba Cancer Research Center, Chaim Sheba Medical Center, 4Department of Pathology, Brigham and Women's Hospital

The fallopian tube (FT) is emerging as an alternative site of origin for serous ovarian carcinoma (SOC). This protocol describes a novel method for the isolation and ex vivo culture of fallopian tube epithelial cells. This system recapitulates the in vivo epithelium and allows the study of SOC pathogenesis.

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Immunology and Infection

Measuring Growth and Gene Expression Dynamics of Tumor-Targeted S. Typhimurium Bacteria
Tal Danino *1, Arthur Prindle *2, Jeff Hasty 2,3,4, Sangeeta Bhatia 1,5,6,7,8
1Health Sciences and Technology, Massachusetts Institute of Technology, 2Department of Bioengineering, University of California, San Diego , 3Biocircuits Institute, University of California, San Diego , 4Molecular Biology Section, Division of Biological Science, University of California, San Diego , 5Broad Institute of Harvard and MIT, 6Department of Medicine, Brigham and Women's Hospital, 7Electrical Engineering and Computer Science and David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, 8Howard Hughes Medical Institute

The goal of these experiments is to generate quantitative time-course data on the growth and gene expression dynamics of attenuated S. typhimurium bacterial colonies growing inside tumors. This video covers tumor cell preparation and implantation, bacteria preparation and injection, whole-animal luminescence imaging, tumor excision, and bacterial colony counting.

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Bioengineering

Systematic Analysis of In Vitro Cell Rolling Using a Multi-well Plate Microfluidic System
Oren Levy 1,2,3,4,5, Priya Anandakumaran 1,2,3,4,5, Jessica Ngai 1,2,3,4,5, Rohit Karnik 6, Jeffrey M. Karp 1,2,3,4,5
1Division of Biomedical Engineering, Department of Medicine, Brigham and Women's Hospital, 2Center for Regenerative Therapeutics, Brigham and Women's Hospital, 3Harvard Medical School, Harvard University, 4Harvard Stem Cell Institute, Harvard University, 5Harvard-MIT Division of Health Sciences and Technology, 6Department of Mechanical Engineering, Massachusetts Institute of Technology

This study used a multi-well plate microfluidic system, significantly increasing throughput of cell rolling studies under physiologically relevant shear flow. Given the importance of cell rolling in the multi-step cell homing cascade and the importance of cell homing following systemic delivery of exogenous populations of cells in patients, this system offers potential as a screening platform to improve cell-based therapy.

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Medicine

Dried Blood Spot Collection of Health Biomarkers to Maximize Participation in Population Studies
Michael W. Ostler 1, James H. Porter 1,2, Orfeu M. Buxton 1,2,3,4
1Center for Population and Development Studies, Harvard School of Public Health, 2Department of Medicine, Brigham and Women's Hospital, 3Division of Sleep Medicine, Harvard Medical School, 4Department of Biobehavioral Health, Pennsylvania State University

Biomarkers are directly-measured biological indicators of disease or health. In population and social sciences, biomarkers need to be easy to obtain, transport, and analyze. Dried Blood Spot (DBS) collection meets this need, can be collected in the field with high response rates and analyzed for a variety of biomarkers.

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Immunology and Infection

Cecal Ligation and Puncture-induced Sepsis as a Model To Study Autophagy in Mice
Ilias I. Siempos 1,3, Hilaire C. Lam 1, Yan Ding 2, Mary E. Choi 2, Augustine M. K. Choi 1, Stefan W. Ryter 1
1Department of Medicine, Division of Pulmonary and Critical Care Medicine, Brigham and Women's Hospital, 2Department of Medicine, Renal Division, Brigham and Women's Hospital, Harvard Medical School, 3First Department of Critical Care Medicine and Pulmonary Services, University of Athens Medical School, Evangelismos Hospital, Athens, Greece

Experimental sepsis can be induced in mice using the cecal ligation and puncture (CLP) method. Current protocols to assess autophagy in vivo in the context of CLP-induced sepsis are presented here: A protocol for measuring autophagy using (GFP)-LC3 mice, and a protocol for measuring autophagosome formation by electron microscopy.

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Medicine

Calcification of Vascular Smooth Muscle Cells and Imaging of Aortic Calcification and Inflammation
Caitlin O'Rourke *1, Georgia Shelton *1,2, Joshua D. Hutcheson 3,4, Megan F. Burke 2, Trejeeve Martyn 1, Timothy E. Thayer 2, Hannah R. Shakartzi 1, Mary D. Buswell 1, Robert E. Tainsh 1, Binglan Yu 1,4, Aranya Bagchi 1,4, David K. Rhee 2,4, Connie Wu 1,2,4, Matthias Derwall 5, Emmanuel S. Buys 1,4, Paul B. Yu 3,4, Kenneth D. Bloch 1,2,4, Elena Aikawa 3,4, Donald B. Bloch 1,5,6, Rajeev Malhotra 2,4
1Anesthesia Center for Critical Care Research of the Department of Anesthesia, Critical Care, and Pain Medicine, Massachusetts General Hospital, 2Cardiovascular Research Center and Cardiology Division of the Department of Medicine, Massachusetts General Hospital, 3Cardiovascular Division, Brigham and Women's Hospital, 4Harvard Medical School, 5Department of Anesthesiology, Uniklinik RWTH Aachen, RWTH Aachen University, 6Center for Immunology and Inflammatory Diseases and the Division of Rheumatology, Allergy, and Immunology of the Department of Medicine, Massachusetts General Hospital

Vascular calcification is an important predictor of and contributor to human cardiovascular disease. This protocol describes methods for inducing calcification of cultured primary vascular smooth muscle cells and for quantifying calcification and macrophage burden in animal aortas using near-infrared fluorescence imaging.

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Immunology and Infection

An All-on-chip Method for Rapid Neutrophil Chemotaxis Analysis Directly from a Drop of Blood
Ke Yang *1,2,3, Jiandong Wu *3,4, Ling Zhu 1, Yong Liu 1, Michael Zhang 5, Francis Lin 3,4,6,7
1Institute of Applied Technology, Hefei Institutes of Physical Science, Chinese Academy of Sciences, 2University of Science and Technology of China, 3Department of Physics and Astronomy, University of Manitoba, 4Department of Biosystems Engineering, University of Manitoba, 5Seven Oaks General Hospital, 6Department of Immunology, University of Manitoba, 7Department of Biological Sciences, University of Manitoba

This article provides the detailed method of performing a rapid neutrophil chemotaxis assay by integrating the on-chip neutrophil isolation from whole blood and the chemotaxis test on a single microfluidic chip.

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Biology

Methods to Classify Cytoplasmic Foci as Mammalian Stress Granules
Anaïs Aulas *1,2, Marta M. Fay *1,2, Witold Szaflarski 1,2,3, Nancy Kedersha 1,2, Paul Anderson 1,2, Pavel Ivanov 1,2,4
1Division of Rheumatology, Immunology, and Allergy, Brigham and Women's Hospital, 2Department of Medicine, Harvard Medical School, 3Department of Histology and Embryology, Poznan University of Medical Sciences, 4The Broad Institute of Harvard and M.I.T.

Stress Granules (SGs) are nonmembranous cytoplasmic structures that form in cells exposed to a variety of stresses. SGs contain mRNAs, RNA-binding proteins, small ribosomal subunits, translation-related factors, and various cell signaling proteins. This protocol describes a workflow that uses several experimental approaches to detect, characterize, and quantify bona fide SGs.

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Cancer Research

Modeling Breast Cancer via an Intraductal Injection of Cre-expressing Adenovirus into the Mouse Mammary Gland
Dongxi Xiang 1,2, Luwei Tao 1,2, Zhe Li 1,2
1Division of Genetics, Department of Medicine, Brigham and Women's Hospital, 2Department of Medicine, Harvard Medical School

The goal of this protocol is to describe a new breast cancer modeling approach based on the intraductal injection of Cre-expressing adenovirus into mouse mammary glands. This approach allows both cell-type- and organ-specific manipulation of oncogenic events in a temporally controlled manner.

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