Aseptic Laboratory Techniques: Plating Methods

Erin R. Sanders¹

¹Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles

JoVE 3064

Preparation, Imaging, and Quantification of Bacterial Surface Motility Assays

Nydia Morales-Soto^1,2, Morgen E. Anyan¹, Anne E. Mattingly¹, Chinedu S. Madukoma¹, Cameron W. Harvey³, Mark Alber³, Eric Déziel⁴, Daniel B. Kearns⁵, Joshua D. Shrout^1,2,6

¹Department of Civil and Environmental Engineering and Earth Sciences, University of Notre Dame, ²Eck Institute for Global Health, University of Notre Dame, ³Department of Applied and Computational Mathematics and Statistics, University of Notre Dame, ⁴INRS-Institut Armand-Frappier, ⁵Department of Biology, Indiana University, ⁶Department of Biological Sciences, University of Notre Dame

JoVE 52338

Techniques for the Evolution of Robust Pentose-fermenting Yeast for Bioconversion of Lignocellulose to Ethanol

Patricia J. Slininger¹, Maureen A. Shea-Andersh¹, Stephanie R. Thompson¹, Bruce S. Dien¹, Cletus P. Kurtzman², Leonardo Da Costa Sousa³, Venkatesh Balan³

¹Bioenergy Research Unit, National Center for Agricultural Utilization Research, ²Mycotoxin Prevention and Applied Microbiology Research Unit, National Center for Agricultural Utilization Research, ³Chemical Engineering and Material Science, Great Lakes Bioenergy Center, Michigan State University

JoVE 54227

Development of a More Sensitive and Specific Chromogenic Agar Medium for the Detection of Vibrio parahaemolyticus and Other Vibrio Species

Marie Yeung¹, Trevor Thorsen¹

¹Biological Sciences Department, California Polytechnic State University

JoVE 54493

Real-time Iontophoresis with Tetramethylammonium to Quantify Volume Fraction and Tortuosity of Brain Extracellular Space

John Odackal*¹, Robert Colbourn*^2,3, Namrita Jain Odackal⁴, Lian Tao⁵, Charles Nicholson⁵, Sabina Hrabetova²

¹Department of Medicine, University of Virginia, ²Department of Cell Biology, SUNY Downstate Medical Center, ³Neural and Behavioral Science Graduate Program, SUNY Downstate Medical Center, ⁴Division of Neonatology, University of Virginia, ⁵Department of Neuroscience and Physiology, New York University School of Medicine,

Induction of Cellular Differentiation and Single Cell Imaging of Vibrio parahaemolyticus Swimmer and Swarmer Cells

Jan Heering¹, Alejandra Alvarado¹, Simon Ringgaard¹

¹Department of Ecophysiology, Max Planck Institute for Terrestrial Microbiology

JoVE 55842

Pre-clinical Evaluation of Tyrosine Kinase Inhibitors for Treatment of Acute Leukemia

Sandra Christoph^1,2, Alisa B. Lee-Sherick¹, Susan Sather¹, Deborah DeRyckere¹, Douglas K. Graham¹

¹Department of Pediatrics, University of Colorado Anschutz Medical Campus, ²Department of Bone Marrow Transplantation, University Hospital of Essen

JoVE 50720

Laboratory Techniques Used to Maintain and Differentiate Biotypes of Vibrio cholerae Clinical and Environmental Isolates

Kyle D. Brumfield*¹, Bailey M. Carignan*¹, Jordan N. Ray¹, Panagiota E. Jumpre¹, Mike S. Son¹

¹Department of Biological Sciences, Plymouth State University

JoVE 55760

ODELAY: A Large-scale Method for Multi-parameter Quantification of Yeast Growth

Thurston Herricks¹, Fred D. Mast^1,2, Song Li¹, John D. Aitchison^1,2

¹Institute for Systems Biology, ²Center for Infectious Disease Research

JoVE 55879

A Robust Pneumonia Model in Immunocompetent Rodents to Evaluate Antibacterial Efficacy against S. pneumoniae, H. influenzae, K. pneumoniae, P. aeruginosa or A. baumannii

Jennifer L. Hoover¹, Thomas F. Lewandowski¹, Cindy L. Mininger¹, Christine M. Singley¹, Scott Sucoloski¹, Stephen Rittenhouse¹

¹Antibacterial Discovery Performance Unit, GlaxoSmithKline

JoVE 55068

Standardized Method for High-throughput Sterilization of Arabidopsis Seeds

Benson E Lindsey III*¹, Luz Rivero*¹, Chistopher S Calhoun¹, Erich Grotewold^1,2, Jelena Brkljacic¹

¹Arabidopsis Biological Resource Center, Center for Applied Plant Sciences, The Ohio State University, ²Department of Molecular Genetics, Center for Applied Plant Sciences, The Ohio State University

Monitoring Spatial Segregation in Surface Colonizing Microbial Populations

Theresa Hölscher¹, Anna Dragoš¹, Ramses Gallegos-Monterrosa¹, Marivic Martin¹, Eisha Mhatre¹, Anne Richter¹, Ákos T. Kovács¹

¹Terrestrial Biofilms Group, Institute of Microbiology, Friedrich Schiller University, Jena

JoVE 54752

A Quantitative Fitness Analysis Workflow

A.P. Banks*¹, C. Lawless*¹, D.A. Lydall¹

¹Institute for Cell and Molecular Biosciences, Newcastle University Medical School

JoVE 4018

Synthesis of Infectious Bacteriophages in an E. coli-based Cell-free Expression System

Mark Rustad¹, Allen Eastlund², Ryan Marshall¹, Paul Jardine², Vincent Noireaux¹

¹School of Physics and Astronomy, University of Minnesota, ²Department of Diagnostic and Biological Sciences and Institute for Molecular Virology, University of Minnesota

Generation of Marked and Markerless Mutants in Model Cyanobacterial Species

David J. Lea-Smith¹, Ravendran Vasudevan¹, Christopher J. Howe¹

¹Department of Biochemistry, University of Cambridge

JoVE 54001

Quantification of Information Encoded by Gene Expression Levels During Lifespan Modulation Under Broad-range Dietary Restriction in C. Elegans

Dhaval S Patel*¹, Giovanni Diana*¹, Eugeni V Entchev¹, Mei Zhan^2,3,4, Hang Lu^2,3,4, QueeLim Ch'ng¹

¹Centre for Developmental Neurobiology, King's College London, ²Interdisciplinary Bioengineering Graduate Program, Georgia Institute of Technology, ³Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, ⁴School of Chemical & Biomolecular Engineering, Georgia Institute of Technology

Monitoring Intraspecies Competition in a Bacterial Cell Population by Cocultivation of Fluorescently Labelled Strains

Lorena Stannek¹, Richard Egelkamp¹, Katrin Gunka¹, Fabian M. Commichau¹

¹Department of General Microbiology, Georg-August University

JoVE 51196

Isolation of Native Soil Microorganisms with Potential for Breaking Down Biodegradable Plastic Mulch Films Used in Agriculture

Graham Bailes¹, Margaret Lind¹, Andrew Ely¹, Marianne Powell², Jennifer Moore-Kucera³, Carol Miles², Debra Inglis², Marion Brodhagen¹

¹Biology Department, Western Washington University, ²Washington State University Northwestern Research and Extension Center, ³Department of Plant and Soil Science, Texas Tech University

JoVE 50373

Methodologies for Studying B. subtilis Biofilms as a Model for Characterizing Small Molecule Biofilm Inhibitors

Tabitha Bucher¹, Elena Kartvelishvily², Ilana Kolodkin-Gal¹

¹Department of Molecular Genetics, Weizmann Institute of Science, ²Electron Microscopy Unit, Weizmann Institute of Science

JoVE 54612

In-vivo Centrifugation of Drosophila Embryos

Susan L. Tran¹, Michael A. Welte¹

¹Department of Biology, University of Rochester

JoVE 2005

Biochemical and High Throughput Microscopic Assessment of Fat Mass in Caenorhabditis Elegans

Elizabeth C. Pino¹, Christopher M. Webster¹, Christopher E. Carr², Alexander A. Soukas¹

¹Center for Human Genetic Research and Department of Medicine, Massachusetts General Hospital and Harvard Medical School, ²Department of Earth, Atmospheric, and Planetary Sciences, Massachusetts Institute of Technology

JoVE 50180

Methods to Inhibit Bacterial Pyomelanin Production and Determine the Corresponding Increase in Sensitivity to Oxidative Stress

Laura M. Ketelboeter¹, Sonia L. Bardy¹

¹Department of Biological Sciences, University of Wisconsin Milwaukee

JoVE 53105

Agar-Block Microcosms for Controlled Plant Tissue Decomposition by Aerobic Fungi

Jonathan S. Schilling¹, K. Brook Jacobson*¹

¹Department of Bioproducts and Biosystems Engineering, University of Minnesota

JoVE 2283

Large-scale Gene Knockdown in C. elegans Using dsRNA Feeding Libraries to Generate Robust Loss-of-function Phenotypes

Kathryn N. Maher¹, Mary Catanese², Daniel L. Chase³

¹Molecular and Cellular Biology Program, University of Massachusetts, Amherst, ²Neuroscience and Behavior Program, University of Massachusetts, Amherst, ³Department of Biochemistry and Molecular Biology, University of Massachusetts, Amherst

JoVE 50693

Methods for Facilitating Microbial Growth on Pulp Mill Waste Streams and Characterization of the Biodegradation Potential of Cultured Microbes

Stephanie L. Mathews^1,2, Ali S. Ayoub², Joel Pawlak², Amy M. Grunden¹

¹Department of Plant and Microbial Biology, North Carolina State University, ²Department of Forest Biomaterials, North Carolina State University

JoVE 51373

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

Chris Troll¹, David Alexander¹, Jennifer Allen¹, Jacob Marquette¹, Manel Camps¹

¹Department of Microbiology & Environmental Toxicology, University of California Santa Cruz - UCSC

JoVE 2505

Using Coculture to Detect Chemically Mediated Interspecies Interactions

Elizabeth Anne Shank¹

¹Department of Biology, University of North Carolina at Chapel Hill

JoVE 50863

Utilizing the Ethylene-releasing Compound, 2-Chloroethylphosphonic Acid, as a Tool to Study Ethylene Response in Bacteria

Richard V. Augimeri¹, Andrew J. Varley¹, Janice L. Strap¹

¹Molecular Microbial Biochemistry Laboratory, Faculty of Science (Applied Bioscience Program), University of Ontario Institute of Technology

JoVE 54682

Aseptic Technique in Environmental Science

JoVE Science Education

Source: Laboratories of Dr. Ian Pepper and Dr. Charles Gerba - Arizona University
Demonstrating Author: Luisa Ikner

Aseptic technique is a fundamental skill widely practiced in the field of environmental microbiology that requires a balance of mindfulness and practice in the laboratory. Proper use of this technique reduces the likelihood of bacterial or fungal contamination of reagents, culture media, and environmental samples. Aseptic technique is also vital to ensure data integrity and maintain the purity of culture libraries that may be comprised of very rare and difficult to culture isolates. Sources of contamination in the laboratory environment include airborne microorganisms (including those adhering to dust and lint particles), microbes present on the laboratory bench workspace or on unsterilized glassware or equipment, and microbes transferred from the body and hair of the researcher. The use of aseptic technique is also a safety measure that lowers the potential for the transmission of microorganisms to researchers, which is particularly important when working with pathogens.…

A Protocol for Phage Display and Affinity Selection Using Recombinant Protein Baits

Rekha Kushwaha¹, Kim R. Schäfermeyer¹, A. Bruce Downie¹

¹Department of Horticulture, University of Kentucky

JoVE 50685

CAPRRESI: Chimera Assembly by Plasmid Recovery and Restriction Enzyme Site Insertion

Orlando Santillán¹, Miguel A. Ramírez-Romero², Guillermo Dávila³

¹Centro de Ciencias Genómicas, Universidad Nacional Autónoma de México, ²Oncomedic Incorporation, Ciudad de México, ³Laboratorio Internacional de Investigación sobre el Genoma Humano, Universidad Nacional Autónoma de México

JoVE 55526

Rapid Identification of Chemical Genetic Interactions in Saccharomyces cerevisiae

David Dilworth¹, Christopher J. Nelson¹

¹Department of Biochemistry and Microbiology, University of Victoria

JoVE 52345

Visualization of the Charcoal Agar Resazurin Assay for Semi-quantitative, Medium-throughput Enumeration of Mycobacteria

Ben Gold¹, Julia Roberts*¹, Yan Ling*¹, Landys Lopez Quezada¹, Jou Glasheen¹, Elaine Ballinger¹, Selin Somersan-Karakaya², Thulasi Warrier¹, Carl Nathan¹

¹Departments of Microbiology & Immunology, Weill Cornell Medical College, ²Medicine, Weill Cornell Medical College

JoVE 54690

Assessing Bacterial Invasion of Cardiac Cells in Culture and Heart Colonization in Infected Mice Using Listeria monocytogenes

P. David McMullen¹, Nancy E. Freitag¹

¹Department of Microbiology and Immunology, College of Medicine, University of Illinois at Chicago

JoVE 52497

The Soft Agar Colony Formation Assay

Stanley Borowicz¹, Michelle Van Scoyk², Sreedevi Avasarala², Manoj Kumar Karuppusamy Rathinam², Jordi Tauler², Rama Kamesh Bikkavilli², Robert A. Winn^2,3

¹Department of Hematology and Oncology, University of Illinois at Chicago, ²Department of Pulmonary, Critical Care, Sleep, and Allergy, University of Illinois at Chicago, ³Jesse Brown Veterans Affairs Medical Center

JoVE 51998

Culturing and Maintaining Clostridium difficile in an Anaerobic Environment

Adrianne N. Edwards¹, Jose M. Suárez¹, Shonna M. McBride¹

¹Department of Microbiology and Immunology, Emory University School of Medicine

JoVE 50787

Generation of Enterobacter sp. YSU Auxotrophs Using Transposon Mutagenesis

Jonathan James Caguiat¹

¹Department of Biological Sciences, Center for Applied Chemical Biology, Youngstown State University

JoVE 51934

Utilization of the Soft Agar Colony Formation Assay to Identify Inhibitors of Tumorigenicity in Breast Cancer Cells

Sachi Horibata¹, Tommy V. Vo², Venkataraman Subramanian³, Paul R. Thompson³, Scott A. Coonrod¹

¹Department of Baker Institute for Animal Health, Cornell University, ²Department of Molecular Biology and Genetics, Cornell University, ³Department of Biochemistry and Molecular Pharmacology, University of Massachusetts Medical School

JoVE 52727

Cultivation of Caenorhabditis elegans in Three Dimensions in the Laboratory

Tong Y. Lee¹, Kyoung-hye Yoon¹, Jin I. Lee¹

¹Division of Biological Science and Technology, Yonsei University

JoVE 55048

Transformation of Probiotic Yeast and Their Recovery from Gastrointestinal Immune Tissues Following Oral Gavage in Mice

Lauren E. Hudson¹, Taryn P. Stewart¹, Milo B. Fasken², Anita H. Corbett², Tracey J. Lamb¹

¹Department of Pediatrics, Emory University School of Medicine, ²Department of Biochemistry, Emory University School of Medicine

JoVE 53453

Expression, Isolation, and Purification of Soluble and Insoluble Biotinylated Proteins for Nerve Tissue Regeneration

Aleesha M. McCormick¹, Natalie A. Jarmusik¹, Elizabeth J. Endrizzi¹, Nic D. Leipzig¹

¹Department of Chemical and Biomolecular Engineering, University of Akron

JoVE 51295

Legionella pneumophila Outer Membrane Vesicles: Isolation and Analysis of Their Pro-inflammatory Potential on Macrophages

Anna Lena Jung¹, Kerstin Hoffmann¹, Christina E. Herkt¹, Christine Schulz¹, Wilhelm Bertrams¹, Bernd Schmeck^1,2,3

¹Institute for Lung Research, Universities of Giessen and Marburg Lung Center, Philipps-University Marburg, ²German Center for Lung Research, ³Department of Medicine, Pulmonary and Critical Care Medicine, University Medical Center Giessen and Marburg

JoVE 55146

3D Whole-heart Myocardial Tissue Analysis

Hans Thijs Van den Broek¹, Leon De Jong^1,2, Pieter A. Doevendans¹, Steven A.J. Chamuleau¹, Frebus J. Van Slochteren*¹, René Van Es*¹

¹Department of Cardiology, Division Heart and Lungs, University Medical Center Utrecht, ²MIRA Institute, University Twente

JoVE 54974

Combination of Adhesive-tape-based Sampling and Fluorescence in situ Hybridization for Rapid Detection of Salmonella on Fresh Produce

Bledar Bisha¹, Byron F. Brehm-Stecher²

¹Center for Meat Safety and Quality, Department of Animal Sciences, Colorado State University, ²Rapid Microbial Detection and Control Laboratory, Department of Food Science and Human Nutrition, Iowa State University

JoVE 2308

Protocols for Implementing an Escherichia coli Based TX-TL Cell-Free Expression System for Synthetic Biology

Zachary Z. Sun*¹, Clarmyra A. Hayes*², Jonghyeon Shin³, Filippo Caschera⁴, Richard M. Murray², Vincent Noireaux⁴

¹Department of Biology, California Institute of Technology, ²Department of Bioengineering, California Institute of Technology, ³Synthetic Biology Center, Department of Bioengineering, Massachusetts Institute of Technology, ⁴School of Physics and Astronomy, University of Minnesota

JoVE 50762

Automated Analysis of a Nematode Population-based Chemosensory Preference Assay

Cynthia M. Chai^1,2, Christopher J. Cronin^1,2, Paul W. Sternberg^1,2

¹Division of Biology and Bioengineering, California Institute of Technology, ²Howard Hughes Medical Institute, California Institute of Technology

Ice-Cap: A Method for Growing Arabidopsis and Tomato Plants in 96-well Plates for High-Throughput Genotyping

Shih-Heng Su¹, Katie A. Clark², Nicole M. Gibbs¹, Susan M. Bush¹, Patrick J. Krysan¹

¹Horticulture Department, University of Wisconsin-Madison, ²Department of Zoology, Oregon State University

JoVE 3280

Modeling Neural Immune Signaling of Episodic and Chronic Migraine Using Spreading Depression In Vitro

Aya D. Pusic*¹, Yelena Y. Grinberg*¹, Heidi M. Mitchell², Richard P. Kraig¹

¹Department of Neurology and Committee on Neurobiology, The University of Chicago Medical Center, ²Department of Neurology, The University of Chicago Medical Center

JoVE 2910

Conjugative Mating Assays for Sequence-specific Analysis of Transfer Proteins Involved in Bacterial Conjugation

Fettah Erdogan¹, Cristina Lento¹, Ayat Yaseen¹, Roksana Nowroozi-Dayeni¹, Sasha Kheyson¹, Gerald F. Audette^1,2

¹Department of Chemistry, York University, ²The Centre for Research on Biomolecular Interactions, York University

JoVE 54854

From a Natural Product to Its Biosynthetic Gene Cluster: A Demonstration Using Polyketomycin from Streptomyces diastatochromogenes Tü6028

Anja Greule¹, Songya Zhang¹, Thomas Paululat², Andreas Bechthold¹

¹Department of Pharmaceutical Biology and Biotechnology, Albert-Ludwigs-Universität Freiburg, Germany, ²Department of Chemistry and Biology, Universität Siegen

JoVE 54952