Harvard Medical School and Boston Children's Hospital

12 articles published in JoVE

• Bioengineering

  
  Modeling Healthy and Dysbiotic Vaginal Microenvironments in a Human Vagina-on-a-Chip Aakanksha Gulati¹, Alicia Jorgenson¹, Abidemi Junaid¹, Donald E. Ingber^{1,2,3 1}Wyss Institute for Biologically Inspired Engineering, Harvard University, ² This article describes a protocol for creating a microfluidic vagina-on-a-chip (Vagina Chip) culture device that enables the study of human host interactions with a living vaginal microbiome under microaerophilic conditions. This chip can be used as a tool to investigate vaginal diseases as well as to develop and test potential therapeutic countermeasures.

• Bioengineering

  
  Scalable Fabrication of Stretchable, Dual Channel, Microfluidic Organ Chips Richard Novak*¹, Meredyth Didier*^1,2, Elizabeth Calamari¹, Carlos F Ng¹, Youngjae Choe¹, Susan L Clauson¹, Bret A Nestor¹, Jefferson Puerta¹, Rachel Fleming¹, Sasan J Firoozinezhad¹, Donald E Ingber^{1,3,4 1}Wyss Institute for Biologically Inspired Engineering, Harvard University, ²Apple, Inc, ³Harvard John A. Paulson School of Engineering and Applied Sciences, Harvard University, ⁴ Here, we present a protocol that describes the fabrication of stretchable, dual channel, organ chip microfluidic cell culture devices for recapitulating organ-level functionality in vitro.

• Immunology and Infection

  
  Deep Dermal Injection As a Model of Candida albicans Skin Infection for Histological Analyses William Santus¹, Francesca Mingozzi¹, Marina Vai¹, Francesca Granucci*¹, Ivan Zanoni*^{1,2 1}Department of Biotechnology and Biosciences, University of Milano-Bicocca, ² Here we describe a protocol that allows histological and molecular analysis of skin samples after Candida albicans intradermal injection. This protocol maintains the structural integrity of the skin and allows for the localization of tissue-resident or newly recruited immune cells as well as the pathogen distribution.

• Genetics

  
  A Fast and Quantitative Method for Post-translational Modification and Variant Enabled Mapping of Peptides to Genomes Christoph N. Schlaffner^1,2,3, Georg J. Pirklbauer², Andreas Bender³, Judith A.J. Steen¹, Jyoti S. Choudhary^{2,4 1} Here we present the proteogenomic tool PoGo and protocols for fast, quantitative, post-translational modification and variant enabled mapping of peptides identified through mass spectrometry onto reference genomes. This tool is of use to integrate and visualize proteogenomic and personal proteomic studies interfacing with orthogonal genomics data.

• Cancer Research

  
  A Time-lapse, Label-free, Quantitative Phase Imaging Study of Dormant and Active Human Cancer Cells Jing Huang*^1,2, Peng Guo*^1,2, Marsha A. Moses^{1,2 1} Dormant and active cancer cell phenotypes were characterized using quantitative phase imaging. Cell proliferation, migration, and morphology assays were integrated and analyzed in one simple method.

• Medicine

  
  Interictal High Frequency Oscillations Detected with Simultaneous Magnetoencephalography and Electroencephalography as Biomarker of Pediatric Epilepsy Christos Papadelis¹, Eleonora Tamilia¹, Steven Stufflebeam², Patricia E. Grant¹, Joseph R. Madsen³, Phillip L. Pearl⁴, Naoaki Tanaka^{2 1} 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.

• Neuroscience

  
  Characterizing Multiscale Mechanical Properties of Brain Tissue Using Atomic Force Microscopy, Impact Indentation, and Rheometry Elizabeth Peruski Canovic¹, Bo Qing², Aleksandar S. Mijailovic³, Anna Jagielska², Matthew J. Whitfield², Elyza Kelly⁴, Daria Turner⁴, Mustafa Sahin⁴, Krystyn J. Van Vliet^{1,2 1}Department of Materials Science and Engineering, Massachusetts Institute of Technology, ²Department of Biological Engineering, Massachusetts Institute of Technology, ³Department of Mechanical Engineering, Massachusetts Institute of Technology, ⁴ We present a set of techniques to characterize the viscoelastic mechanical properties of brain at the micro-, meso-, and macro-scales.

• Bioengineering

  
  Co-culture of Living Microbiome with Microengineered Human Intestinal Villi in a Gut-on-a-Chip Microfluidic Device Hyun Jung Kim¹, Jaewon Lee¹, Jin-Ha Choi¹, Anthony Bahinski², Donald E. Ingber^{2,3,4 1}Department of Biomedical Engineering, The University of Texas at Austin, ²Wyss Institute for Biologically Inspired Engineering at Harvard University, ³ We describe an in vitro protocol to co-culture gut microbiome and intestinal villi for an extended period using a human gut-on-a-chip microphysiological system.

• Biology

  
  Rapid Isolation And Purification Of Mitochondria For Transplantation By Tissue Dissociation And Differential Filtration Janine M. Preble¹, Christina A. Pacak², Hiroshi Kondo¹, Allison A. MacKay², Douglas B. Cowan², James D. McCully^{1 1}Division of Cardiothoracic Surgery, Beth Israel Deaconess Medical Center and Harvard Medical School, ² A method for rapid isolation of mitochondria from mammalian tissue biopsies is described. Rat liver or skeletal muscle preparations were homogenized with a commercial tissue dissociator and mitochondria were isolated by differential filtration through nylon mesh filters. Mitochondrial isolation time is <30 min compared to 60 - 100 min using alternative methods.

• Biology

  
  Tissue Triage and Freezing for Models of Skeletal Muscle Disease Hui Meng¹, Paul M.L. Janssen², Robert W. Grange³, Lin Yang⁴, Alan H. Beggs⁵, Lindsay C. Swanson⁵, Stacy A. Cossette^1,6, Alison Frase⁷, Martin K. Childers⁸, Henk Granzier⁹, Emanuela Gussoni⁵, Michael W. Lawlor^{1 1}Division of Pediatric Pathology, Department of Pathology and Laboratory Medicine, Medical College of Wisconsin, ²Department of Physiology and Cell Biology, The Ohio State University, ³Department of Human Nutrition, Foods and Exercise, Virginia Tech, ⁴Division of Biomedical Informatics, Department of Biostatistics, Department of Computer Science, University of Kentucky, ⁵ The analysis of skeletal muscle tissues to determine structural, functional, and biochemical properties is greatly facilitated by appropriate preparation. This protocol describes appropriate methods to prepare skeletal muscle tissue for a broad range of phenotyping studies.

• Neuroscience

  
  An Engulfment Assay: A Protocol to Assess Interactions Between CNS Phagocytes and Neurons Dorothy P. Schafer¹, Emily K. Lehrman¹, Christopher T. Heller¹, Beth Stevens^{1 1} Microglia are the resident immune cells of the central nervous system (CNS) with a high capacity to phagocytose or engulf material in their extracellular environment. Here, a broadly applicable, reliable, and highly quantitative assay for visualizing and measuring microglia-mediated engulfment of synaptic components is described.

• Biology

  
  Intraductal Injection for Localized Drug Delivery to the Mouse Mammary Gland Silva Krause¹, Amy Brock², Donald E. Ingber^{1,2,3 1} A protocol for the non-invasive intraductal delivery of aqueous reagents to the mouse mammary gland is described. The method takes advantage of localized injection into the nipples of mammary glands targeting mammary ducts specifically. This technique is adaptable for a variety of compounds including siRNA, chemotherapeutic agents and small molecules.