Protocol
Live-imaging of the Drosophila Pupal Eye
Mark B. Hellerman, Richard H. Choe, Ruth I. Johnson
Biology Department, Wesleyan University
This protocol presents an efficient method for imaging the live Drosophila pupal eye neuroepithelium. This method compensates for tissue movement and uneven topology, enhances visualization of cell boundaries through the use of multiple GFP-tagged junction proteins, and uses an easily-assembled imaging rig.
State of the Art Cranial Ultrasound Imaging in Neonates
Ginette M. Ecury-Goossen1, Fleur A. Camfferman2, Lara M. Leijser3,4, Paul Govaert1,5, Jeroen Dudink1,2
1Department of Pediatrics, Division of Neonatology, Erasmus MC-Sophia Children's Hospital, 2Department of Radiology, Erasmus MC-Sophia Children's Hospital, 3Department of Pediatrics, Division of Neonatology, UZ Brussel, 4Department of Pediatrics, Division of Neonatology, Leiden University Medical Center, 5Department of Pediatrics, Division of Neonatology, Isala Hospital, 6Department of Pediatrics, Koningin Paola Children's Hospital
Cranial ultrasound (CUS) is a valuable tool for brain imaging in critically ill neonates. This video shows a comprehensive approach for neonatal (Doppler) CUS for both clinical and research purposes, including a bedside demonstration of the technique.
A Technical Perspective in Modern Tree-ring Research - How to Overcome Dendroecological and Wood Anatomical Challenges
Holger Gärtner, Paolo Cherubini, Patrick Fonti, Georg von Arx, Loïc Schneider, Daniel Nievergelt, Anne Verstege, Alexander Bast, Fritz H. Schweingruber, Ulf Büntgen
Landscape Dynamics / Dendroecology, Swiss Federal Research Institute WSL
Here we present a protocol outlining how to sample wooden specimens for the overall assessment of their growth structures. Macro- and microscopic preparation and visualization techniques necessary to generate well-replicated and highly resolved wood anatomical and dendroecological dataset, are described are described.
Preparation, Imaging, and Quantification of Bacterial Surface Motility Assays
Nydia Morales-Soto1,2, Morgen E. Anyan1, Anne E. Mattingly1, Chinedu S. Madukoma1, Cameron W. Harvey3, Mark Alber3, Eric Déziel4, Daniel B. Kearns5, Joshua D. Shrout1,2,6
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
Swarming motility is influenced by physical and environmental factors. We describe a two-phase protocol and guidelines to circumvent the challenges commonly associated with swarm assay preparation and data collection. A macroscopic imaging technique is employed to obtain detailed information on swarm behavior that is not provided by current analysis techniques.
Self-assembly of Complex Two-dimensional Shapes from Single-stranded DNA Tiles
Bryan Wei1, Michelle K. Vhudzijena2, Joanna Robaszewski2, Peng Yin2,3
1Tsinghua-Peking Center for Life Sciences, School of Life Sciences, Tsinghua University, 2Wyss Institute for Biologically Inspired Engineering, Harvard University, 3Department of Systems Biology, Harvard Medical School
DNA tiling is an effective approach to make programmable nanostructures. We describe the protocols to construct complex two-dimensional shapes by the self-assembly of single-stranded DNA tiles.
Quantifying Learning in Young Infants: Tracking Leg Actions During a Discovery-learning Task
Barbara Sargent1, Hendrik Reimann2, Masayoshi Kubo3, Linda Fetters1
1Division of Biokinesiology & Physical Therapy at the Herman Ostrow School of Dentistry, University of Southern California, 2Department of Kinesiology, Temple University, 3Department of Physical Therapy, Niigata University of Health and Welfare
A method is described in which 3-4 month old infants learn a task by discovery and their leg movements are captured to quantify the learning process.
Therapeutic Effectiveness of a Dietary Supplement for Management of Halitosis in Dogs
Alessandro Di Cerbo1, Federica Pezzuto2, Sergio Canello3, Gianandrea Guidetti3, Beniamino Palmieri1,4
1Department of Surgery and Surgical Specialties, Azienda Ospedaliero-Universitaria Policlinico di Modena, 2Department of Clinical Microbiology, Universtity of Modena and Reggio Emilia, 3Sanypet S.p.a, 4AIRMO Center Milan
We describe a simple approach for diagnosis of halitosis in dogs as well as a dietary approach for its management. This protocol may be extended to the management of halitosis in humans in the near future.
Epithelial Cell Repopulation and Preparation of Rodent Extracellular Matrix Scaffolds for Renal Tissue Development
Joseph S. Uzarski1,2, Jimmy Su1,2,3,4, Yan Xie1,2, Zheng J. Zhang1,2, Heather H. Ward5, Angela Wandinger-Ness6, William M. Miller7,8, Jason A. Wertheim1,2,3,4,8,9
1Comprehensive Transplant Center, Feinberg School of Medicine, Northwestern University, 2Department of Surgery, Feinberg School of Medicine, Northwestern University, 3Department of Biomedical Engineering, Northwestern University, 4Simpson Querrey Institute for BioNanotechnology in Medicine, Northwestern University, 5Department of Internal Medicine, University of New Mexico HSC, 6Department of Pathology, University of New Mexico HSC, 7Department of Chemical and Biological Engineering, Northwestern University, 8Chemistry of Life Processes Institute, Northwestern University, 9Department of Surgery, Jesse Brown VA Medical Center
This protocol describes decellularization of Sprague Dawley rat kidneys by antegrade perfusion of detergents through the vasculature, producing acellular renal extracellular matrices that serve as templates for repopulation with human renal epithelial cells. Recellularization and use of the resazurin perfusion assay to monitor growth is performed within specially-designed perfusion bioreactors.
Measuring Rates of Herbicide Metabolism in Dicot Weeds with an Excised Leaf Assay
Rong Ma, Joshua J. Skelton, Dean E. Riechers
Department of Crop Sciences, University of Illinois
This manuscript describes how herbicide metabolism rates can be effectively quantified with excised leaves from a dicot weed, thereby reducing variability and removing any possible confounding effects of herbicide uptake or translocation typically observed in whole-plant assays.
Interfacing 3D Engineered Neuronal Cultures to Micro-Electrode Arrays: An Innovative In Vitro Experimental Model
Mariateresa Tedesco1, Monica Frega1,2, Sergio Martinoia1, Mattia Pesce3, Paolo Massobrio1
1Department of Informatics, Bioengineering, Robotics and System Engineering (DIBRIS), University of Genova, 2Donders Institute for Brain, Cognition and Behaviour, Department of Cognitive Neuroscience, Radboud University Medical Center, 3Fondazione Istituto Italiano di Tecnologia (IIT)
In this work, a novel experimental model in which 3D neuronal cultures are coupled to planar Micro-Electrode Arrays (MEAs) is presented. 3D networks are built by seeding neurons in a scaffold made up of glass microbeads on which neurons grow and form interconnected 3D structures.
A Structured Rehabilitation Protocol for Improved Multifunctional Prosthetic Control: A Case Study
Aidan Dominic Roche1,2, Ivan Vujaklija3,4, Sebastian Amsüss3,4, Agnes Sturma1,5, Peter Göbel6, Dario Farina3,4, Oskar C. Aszmann1,2
1Christian Doppler Laboratory for Restoration of Extremity Function, 2Department of Surgery, Division of Plastic and Reconstructive Surgery, Medical University of Vienna, 3Department of Neurorehabilitation Engineering, Bernstein Focus Neurotechnology Göttingen, 4University Medical Center Göttingen, Georg-August University, 5University of Applied Sciences FH Campus Wien, 6Research & Development, Otto Bock Healthcare Products GmbH
As prosthetic development moves towards the goal of natural control, harnessing amputees’ inherent ability to learn new motor skills may enable proficiency. This manuscript describes a structured rehabilitation protocol, which includes imitation, repetition, and reinforcement learning strategies, for improved multifunctional prosthetic control.
Nasal Wipes for Influenza A Virus Detection and Isolation from Swine
Jacqueline M. Nolting, Christine M. Szablewski, Jody L. Edwards, Sarah W. Nelson, Andrew S. Bowman
Department of Veterinary Preventive Medicine, The Ohio State University
The authors present a protocol to collect swine nasal wipes to detect and isolate influenza A viruses.
Disclosures
No conflicts of interest declared.
