Worcester Polytechnic Institute View Institution's Website 15 articles published in JoVE Cancer Research Shear Assay Protocol for the Determination of Single-Cell Material Properties Luke J. Holen*1, Killian Onwudiwe*1, Julian Najera1, Maksym Zarodniuk1, John D. Obayemi2, Winston O. Soboyejo2, Meenal Datta1 1Department of Aerospace and Mechanical Engineering, University of Notre Dame, 2Departments of Mechanical and Biomedical Engineering, Worcester Polytechnic Institute This protocol outlines the quantification of the mechanical properties of cancerous and non-cancerous cell lines in vitro. Conserved differences in the mechanics of cancerous and normal cells can act as a biomarker that may have implications in prognosis and diagnosis. Neuroscience Automated Multimodal Stimulation and Simultaneous Neuronal Recording from Multiple Small Organisms Hamilton White1,2, Vanessa Kamara1, Veronika Gorski1, Molly Busby1, Dirk R. Albrecht1,3 1Department of Biomedical Engineering, Worcester Polytechnic Institute, 2Graduate School of Biomedical Sciences, UMass Chan Medical School, 3Department of Biology and Biotechnology, Worcester Polytechnic Institute We present a method for the flexible chemical and multimodal stimulation and recording of simultaneous neural activity from many Caenorhabditis elegans worms. This method uses microfluidics, open-source hardware and software, and supervised automated data analysis to enable the measurement of neuronal phenomena such as adaptation, temporal inhibition, and stimulus crosstalk. Bioengineering Assembly of Cell Mimicking Supported and Suspended Lipid Bilayer Models for the Study of Molecular Interactions Christina M. Bailey-Hytholt1, Veronica LaMastro2, Anita Shukla2 1Department of Chemical Engineering, Worcester Polytechnic Institute, 2Center for Biomedical Engineering, School of Engineering, Brown University This protocol describes the formation of cell mimicking uni-lipid and multi-lipid vesicles, supported lipid bilayers, and suspended lipid bilayers. These in vitro models can be adapted to incorporate a variety of lipid types and can be used to investigate various molecule and macromolecule interactions. Engineering Controlling Flow Speeds of Microtubule-Based 3D Active Fluids Using Temperature Teagan E. Bate1, Edward J. Jarvis1, Megan E. Varney1, Kun-Ta Wu1,2 1Department of Physics, Worcester Polytechnic Institute, 2Department of Physics, Brandeis University The goal of this protocol is to use temperature to control the flow speeds of three-dimensional active fluids. The advantage of this method not only allows for regulating flow speeds in situ but also enables dynamic control, such as periodically tuning flow speeds up and down. Biology Live Cell Imaging to Assess the Dynamics of Metaphase Timing and Cell Fate Following Mitotic Spindle Perturbations Dayna L. Mercadante*1, Elizabeth A. Crowley*1, Amity L. Manning1 1Department of Biology and Biotechnology, Worcester Polytechnic Institute Here we present a protocol to assess the dynamics of spindle formation and mitotic progression. Our application of time-lapse imaging enables the user to identify cells at various stages of mitosis, track and identify mitotic defects, and analyze spindle dynamics and mitotic cell fate upon exposure to anti-mitotic drugs. Bioengineering Two Methods for Decellularization of Plant Tissues for Tissue Engineering Applications Michal Adamski1, Gianluca Fontana2, Joshua R. Gershlak4, Glenn R. Gaudette4, Hau D. Le1, William L. Murphy2,3 1Department of Surgery, University of Wisconsin-Madison, 2Department of Orthopedics and Rehabilitation, University of Wisconsin School of Medicine and Public Health, 3Department of Biomedical Engineering, University of Wisconsin College of Engineering, 4Department of Biomedical Engineering, Worcester Polytechnic Institute Here we present, and contrast two protocols used to decellularize plant tissues: a detergent-based approach and a detergent-free approach. Both methods leave behind the extracellular matrix of the plant tissues used, which can then be utilized as scaffolds for tissue engineering applications. Bioengineering Fabrication of Custom Agarose Wells for Cell Seeding and Tissue Ring Self-assembly Using 3D-Printed Molds Hannah A. Strobel1, Elizabeth L. Calamari1, Brittany Alphonse1, Tracy A. Hookway1,2, Marsha W. Rolle1 1Biomedical Engineering, Worcester Polytechnic Institute, 2Gladstone Institute for Cardiovascular Disease This protocol describes a platform for fabricating self-assembled tissue rings in variable sizes using a customized 3D-printed plastic mold. PDMS negatives are cured in the 3D-printed mold; then agarose is cast in the cured PDMS negatives. Cells are seeded into the resulting agarose wells where they aggregate into tissue rings. Genetics The Nematode Caenorhabditis Elegans - A Versatile In Vivo Model to Study Host-microbe Interactions Luca Issi1, Meredith Rioux1, Reeta Rao1 1Biology and Biotechnology, Worcester Polytechnic Institute Here, we present the nematode Caenorhabditis elegans as a versatile host model to study microbial interaction. Neuroscience Using an Adapted Microfluidic Olfactory Chip for the Imaging of Neuronal Activity in Response to Pheromones in Male C. Elegans Head Neurons Douglas K. Reilly1, Daniel E. Lawler2, Dirk R. Albrecht1,2, Jagan Srinivasan1 1Department of Biology and Biotechnology, Worcester Polytechnic Institute, 2Department of Biomedical Engineering, Worcester Polytechnic Institute The use of an adapted "olfactory chip" for the efficient calcium imaging of C. elegans males is described here. Studies of male exposure to glycerol and a pheromone are also shown. Immunology and Infection Isolation of the Side Population in Myc-induced T-cell Acute Lymphoblastic Leukemia in Zebrafish Margaret M. Pruitt1,2, Wilfredo Marin1, Michael R. Waarts1, Jill L. O. de Jong1 1Department of Pediatrics, Section of Hematology-Oncology, The University of Chicago, 2Department of Biology and Biotechnology, Worcester Polytechnic Institute Here, we describe a technique to isolate the side population cells from a zebrafish model of myc-induced T-cell acute lymphoblastic leukemia (T-ALL). This side population assay is highly sensitive and is described for zebrafish T-ALL, but it may be applicable to other malignant and non-malignant zebrafish cell types. Bioengineering Measuring the Mechanical Properties of Living Cells Using Atomic Force Microscopy Gawain Thomas1, Nancy A. Burnham1, Terri Anne Camesano2, Qi Wen1 1Department of Physics, Worcester Polytechnic Institute, 2Department of Chemical Engineering, Worcester Polytechnic Institute This paper demonstrates a protocol to characterize the mechanical properties of living cells by means of microindentation using an Atomic Force Microscope (AFM). Bioengineering Directed Cellular Self-Assembly to Fabricate Cell-Derived Tissue Rings for Biomechanical Analysis and Tissue Engineering Tracy A. Gwyther1, Jason Z. Hu1, Kristen L. Billiar1, Marsha W. Rolle1 1Biomedical Engineering Department, Worcester Polytechnic Institute This article outlines a versatile method to create cell-derived tissue rings by cellular self-assembly. Smooth muscle cells seeded into ring-shaped agarose wells aggregate and contract to form robust three-dimensional (3D) tissues within 7 days. Millimeter-scale tissue rings are conducive to mechanical testing and serve as building blocks for tissue assembly. Biology Examining the Conformational Dynamics of Membrane Proteins in situ with Site-directed Fluorescence Labeling Ryan Richards1, Robert E. Dempski1 1Department of Chemistry and Biochemistry, Worcester Polytechnic Institute We will describe a method which measures the kinetics of ion transport of membrane proteins alongside site-specific analysis of conformational changes using fluorescence on single cells. This technique is adaptable to ion channels, transporters and ion pumps and can be utilized to determine distance constraints between protein subunits. Biology Efficient Polyethylene Glycol (PEG) Mediated Transformation of the Moss Physcomitrella patens Yen-Chun Liu1, Luis Vidali1 1Department of Biology and Biotechnology, Worcester Polytechnic Institute- WPI A simple and efficient method to transform Physcomitrella pantens protoplasts is described. This method is adapted from protocols for Physocmitrella protonemal protoplast and Arabidopsis mesophyll protoplast transformation1. Biology Heterokaryon Technique for Analysis of Cell Type-specific Localization Roseann Gammal*1, Krista Baker*1, Destin Heilman1 1Department of Chemistry and Biochemistry, Worcester Polytechnic Institute- WPI A flexible and efficient method for the characterization of cell type-specific protein localization and nucleocytoplasmic shuttling is described. This heterokaryon approach uses fluorescently-labeled fusion proteins to image protein localizations after cell fusion. The protocol is amenable to steady-state localizations or more dynamic determinations based on live cell imaging.
