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  JoVE Developmental Biology


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October, 2006
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Robotics: The application of electronic, computerized control systems to mechanical devices designed to perform human functions. Formerly restricted to industry, but nowadays applied to artificial organs controlled by bionic (bioelectronic) devices, like automated insulin pumps and other prostheses.
 JoVE Biology

Robotics and Dynamic Image Analysis for Studies of Gene Expression in Plant Tissues

1Department of Horticulture and Crop Science, The Ohio State University, 2Department of Plant Pathology, North Carolina State University

JoVE 1733

We report a method for introduction, tracking and quantitative analysis of GFP expression in plant cells. This method utilizes a custom-designed robotics system for semi-continuous image collection from large numbers of samples, over time. We also demonstrate the use of ImageJ and ImageReady for analysis of image series.

 JoVE Bioengineering

High-throughput, Automated Extraction of DNA and RNA from Clinical Samples using TruTip Technology on Common Liquid Handling Robots

1Application Development, Akonni Biosystems, Inc., 2Manufacturing, Akonni Biosystems, Inc., 3Engineering, Akonni Biosystems, Inc., 4Research & Development, Akonni Biosystems, Inc.

JoVE 50356

TruTip is a simple nucleic acid extraction technology whereby a porous, monolithic binding matrix is inserted into a pipette tip. Consequently, the sample preparation format is compatible with most liquid handling instruments, and can be used for many medium to high-throughput clinical applications and sample types.

 JoVE Biology

Genome-wide Protein-protein Interaction Screening by Protein-fragment Complementation Assay (PCA) in Living Cells

1Département de Biologie, Institut de biologie intégrative et des systémes & PROTEO, Université Laval

JoVE 52255

Proteins interact with each other and these interactions determine in a large part their functions. Protein interaction partners can be identified at high-throughput in vivo using a yeast fitness assay based on the dihydrofolate reductase protein-fragment complementation assay (DHFR-PCA).

 JoVE Bioengineering

Haptic/Graphic Rehabilitation: Integrating a Robot into a Virtual Environment Library and Applying it to Stroke Therapy

1Department of Bioengineering, University of Illinois at Chicago and Rehabilitation Institute of Chicago, 2Sensory Motor Performance Program, Rehabilitation Institute of Chicago

JoVE 3007

Recently, a vast amount of prospects have come available for human-robot interactive systems. In this paper we outline the integration of a new robotic device with open source software that can rapidly make possible a library of interactive functionality. We then outline a clinical application for a neurorehabilitation application.

 JoVE Biology

A Quantitative Fitness Analysis Workflow

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

JoVE 4018

Quantitative Fitness Analysis (QFA) is a complementary series of experimental and computational methods for estimating microbial culture fitnesses. QFA estimates the effect of genetic mutations, drugs or other applied treatments on microbe growth. Experiments scaling from focussed analysis of single cultures to thousands of parallel cultures can be designed.

 JoVE Biology

Improving the Success Rate of Protein Crystallization by Random Microseed Matrix Screening

1School of Biochemistry, University of Bristol, 2Douglas Instruments

JoVE 50548

Here we describe a general method for random microseed matrix screening. This technique is shown to significantly increase the success rate of protein crystallization screening experiments, reduce the need for optimization, and provide a reliable supply of crystals for data collection and ligand-soaking experiments.

 JoVE Immunology and Infection

High-throughput Quantitative Real-time RT-PCR Assay for Determining Expression Profiles of Types I and III Interferon Subtypes

1Center for Biologics Evaluation and Research, US Food and Drug Administration, 2Center for Drug Evaluation and Research, US Food and Drug Administration

JoVE 52650

This protocol describes a high-throughput qRT-PCR assay for the analysis of type I and III IFN expression signatures. The assay discriminates single base pair differences between the highly similar transcripts of these genes. Through batch assembly and robotic pipetting, the assays are consistent and reproducible.

 JoVE Neuroscience

Using Insect Electroantennogram Sensors on Autonomous Robots for Olfactory Searches

1UMR 7503, Laboratoire Lorrain de Recherche en Informatique et ses Applications (LORIA), Centre National de la Recherche Scientifique (CNRS), 2UMR 1392 iEES-Paris, Institut d'Ecologie et des Sciences de l'Environnement de Paris, 3Physics of Biological Systems, Institut Pasteur

JoVE 51704

We describe a protocol for using insect antennae in the form of electroantennograms (EAGs) on autonomous robots. Our experimental design allows stable recordings within a day and resolves individual odor patches up to 10 Hz. The efficiency of EAG sensors for olfactory searches is demonstrated in driving a robot toward an odor source.

 JoVE Biology

The MultiBac Protein Complex Production Platform at the EMBL

1EMBL Grenoble Outstation and Unit of Virus Host Cell Interactions (UVHCI) UMR5322

JoVE 50159

Protein complexes catalyze key cellular functions. Detailed functional and structural characterization of many essential complexes requires recombinant production. MultiBac is a baculovirus/insect cell system particularly tailored for expressing eukaryotic proteins and their complexes. MultiBac was implemented as an open-access platform, and standard operating procedures developed to maximize its utility.

 JoVE Neuroscience

MPI CyberMotion Simulator: Implementation of a Novel Motion Simulator to Investigate Multisensory Path Integration in Three Dimensions

1Department of Human Perception, Cognition and Action, Max Planck Institute for Biological Cybernetics, 2Laboratoire de Physiologie de la Perception et de l'Action, Collège de France - CNRS, 3Department of Brain and Cognitive Engineering, Korea University

JoVE 3436

An efficient way to gain insight into how humans navigate themselves in three dimensions is described. The method takes advantage of a motion simulator capable of moving observers in ways unattainable by traditional simulators. Results confirm that movement in the horizontal plane is underestimated, while vertical movement is overestimated.

 JoVE Biology

High-throughput Crystallization of Membrane Proteins Using the Lipidic Bicelle Method

1UCLA-DOE Institute for Genomics and Proteomics, University of California Los Angeles, 2Department of Physiology, David Geffen School of Medicine, UCLA

JoVE 3383

Bicelles are lipid/amphiphile mixtures that maintain membrane proteins (MPs) within a lipid bilayer but have unique phase behavior that facilitates high-throughput screening by crystallization robots. This technique has successfully produced a number of high-resolution structures from both prokaryotic and eukaryotic sources. This video describes protocols for generating the lipidic bicelle mixture, incorporating MPs into the bicelle mixture, setting up crystallizations trials (manually as well as robotically) and harvesting crystals from the medium.

 JoVE Immunology and Infection

Scalable High Throughput Selection From Phage-displayed Synthetic Antibody Libraries

1The Recombinant Antibody Network, 2The Banting and Best Department of Medical Research, University of Toronto, 3Antibiome Center, University of California, San Francisco at Mission Bay, 4Department of Biochemistry and Molecular Biology, The University of Chicago

JoVE 51492

A method is described with visual accompaniment for conducting scalable, high throughput selections from phage-displayed combinatorial synthetic antibody libraries against hundreds of antigens simultaneously. Using this parallel approach, we have isolated antibody fragments that exhibit high affinity and specificity for diverse antigens that are functional in standard immunoassays.

 JoVE Medicine

Robotic Ablation of Atrial Fibrillation

1Department of Cardiology, Charité — Universitätsmedizin Berlin, Campus Virchow, 2Department of Cardiology, University Hospital Zurich

JoVE 52560

Pulmonary vein isolation (PVI) with an ablation catheter is a curative treatment for atrial fibrillation (AF). Robotic catheter systems aim to improve catheter steerability. Here, a procedure with a new robotic catheter system is presented. The goal of the procedure is electrical block between pulmonary vein and left atrium.

 JoVE Neuroscience

One Dimensional Turing-Like Handshake Test for Motor Intelligence

1Biomedical Engineering, Ben-Gurion University

JoVE 2492

We present a Turing-like Handshake test administered through a telerobotic system in which the interrogator is holding a robotic stylus and interacting with another party (human or artificial). We use a forced choice method, and extract a measure for the similarity of the artificial model to a human handshake.

 JoVE Chemistry

Origami Inspired Self-assembly of Patterned and Reconfigurable Particles

1Department of Chemical and Biomolecular Engineering, The Johns Hopkins University, 2Department of Chemistry, The Johns Hopkins University

JoVE 50022

We describe experimental details of the synthesis of patterned and reconfigurable particles from two dimensional (2D) precursors. This methodology can be used to create particles in a variety of shapes including polyhedra and grasping devices at length scales ranging from the micro to centimeter scale.

 JoVE Biology

High-throughput Functional Screening using a Homemade Dual-glow Luciferase Assay

1Department of Neurology, Massachusetts General Hospital

JoVE 50282

We present a rapid and inexpensive screening method for identifying transcriptional regulators using high-throughput robotic transfections and a homemade dual-glow luciferase assay. This protocol rapidly generates direct side-by-side functional data for thousands of genes and is easily modifiable to target any gene of interest.

 JoVE Bioengineering

Adaptation of a Haptic Robot in a 3T fMRI

1Institute for Neural Computation, University of California, 2Department of Radiology, University of California, 3Department of Cognitive Science and Program in Neurosciences, University of California

JoVE 3364

The adaptation and use of a haptic robot in a 3T fMRI is described.

 JoVE Bioengineering

High-throughput Protein Expression Generator Using a Microfluidic Platform

1The Mina & Everard Goodman Faculty of Life Sciences, The Nanotechnology Institute, Bar-Ilan University

JoVE 3849

We present a microfluidic approach for the expression of protein arrays. The device consists of thousands of reaction chambers controlled by micro-mechanical valves. The microfluidic device is mated to a microarray-printed gene library. These genes are then transcribed and translated on-chip, resulting in a protein array ready for experimental use.

 JoVE Biology

Array Comparative Genomic Hybridization (Array CGH) for Detection of Genomic Copy Number Variants

1Cytogenetics Department, Guy's & St Thomas' NHS Foundation Trust, 2Cytogenetics Department, Viapath Analytics

JoVE 51718

Array CGH for the detection of genomic copy number variants has replaced G-banded karyotype analysis. This paper describes the technology and its application in a diagnostic service laboratory.

 JoVE Developmental Biology

Scalable 96-well Plate Based iPSC Culture and Production Using a Robotic Liquid Handling System

1InvivoSciences, Inc., 2Gilson, Inc.

JoVE 52755

The protocols described allow laboratories to perform scalable, adherent stem cell culture in high throughput with minimal labor, experience and equipment investment cost using a programmable liquid handling robot and 96-well plates. iPSCs passaged more than 20 times on this system maintained pluripotency, normal karyotypes and differentiated into cardiomyocytes.

 JoVE Behavior

Perceptual and Category Processing of the Uncanny Valley Hypothesis' Dimension of Human Likeness: Some Methodological Issues

1Department of Neuropsychology, University of Zurich

JoVE 4375

Investigation of the Uncanny Valley Hypothesis and affective experience requires an understanding of the hypothesis' dimension of human likeness (DHL). This protocol allows representation of the DHL and examination of categorical perception. Use of the same stimuli and fMRI to distinguish brain regions responsive to physical and category change is illustrated.

 JoVE Bioengineering

Drug-induced Sensitization of Adenylyl Cyclase: Assay Streamlining and Miniaturization for Small Molecule and siRNA Screening Applications

1Department of Medicinal Chemistry and Molecular Pharmacology, Purdue University, 2Quantitative Biology, Eli Lilly and Company

JoVE 51218

Persistent activation of inhibitory G protein-coupled receptors results in sensitization of adenylyl cyclase signaling. To identify the essential molecular pathways, nonbiased approaches are necessary; however, this strategy requires the development of a scalable cell-based cAMP sensitization assay. Herein, we describe a sensitization assay for small molecule and siRNA screening.

 JoVE Biology

Genetic Barcoding with Fluorescent Proteins for Multiplexed Applications

1Department of Biology, San Diego State University

JoVE 52452

Since the discovery of the green fluorescent protein gene, fluorescent proteins have impacted molecular cell biology. This protocol describes how expression of distinct fluorescent proteins through genetic engineering is used for barcoding individual cells. The procedure enables tracking distinct populations in a cell mixture, which is ideal for multiplexed applications.

 JoVE Bioengineering

Fabricating Complex Culture Substrates Using Robotic Microcontact Printing (R-µCP) and Sequential Nucleophilic Substitution

1Department of Biomedical Engineering, University of Wisconsin, Madison, 2Department of Mechanical Engineering, University of Wisconsin, Madison

JoVE 52186

Cell culture substrates functionalized with microscale patterns of biological ligands have immense utility in the field of tissue engineering. Here, we demonstrate the versatile and automated manufacture of tissue culture substrates with multiple, micropatterned poly(ethylene glycol) brushes presenting orthogonal chemistries that enable spatially precise and site-specific immobilization of biological ligands.

 JoVE Bioengineering

Engineering Platform and Experimental Protocol for Design and Evaluation of a Neurally-controlled Powered Transfemoral Prosthesis

1Joint Department of Biomedical Engineering, North Carolina State University & University of North Carolina at Chapel Hill, 2Department of Physical Medicine and Rehabilitation, University of North Carolina School of Medicine, 3Atlantic Prosthetics & Orthotics, LLC

JoVE 51059

Neural-machine interfaces (NMI) have been developed to identify the user's locomotion mode. These NMIs are potentially useful for neural control of powered artificial legs, but have not been fully demonstrated. This paper presented (1) our designed engineering platform for easy implementation and development of neural control for powered lower limb prostheses and (2) an experimental setup and protocol in a laboratory environment to evaluate neurally-controlled artificial legs on patients with lower limb amputations safely and efficiently.

 JoVE Bioengineering

Robotic Production of Cancer Cell Spheroids with an Aqueous Two-phase System for Drug Testing

1Department of Biomedical Engineering, The University of Akron

JoVE 52754

A protocol for robotic printing of cancer cell spheroids in a high throughput 96-well plate format using an aqueous two-phase system is presented.

 JoVE Neuroscience

Closed-loop Neuro-robotic Experiments to Test Computational Properties of Neuronal Networks

1Neuroscience and Brain Technologies, Istituto Italiano di Tecnologia

JoVE 52341

In this paper, an experimental framework to perform closed-loop experiments is presented, in which information processing (i.e., coding and decoding) and learning of neuronal assemblies are studied during the continuous interaction with a robotic body.

 JoVE Immunology and Infection

Establishment and Optimization of a High Throughput Setup to Study Staphylococcus epidermidis and Mycobacterium marinum Infection as a Model for Drug Discovery

1Institute of Biology, Leiden University, 2ZF-screens BV, 3Life Science Methods BV

JoVE 51649

This video article describes the high throughput pipeline that has been successfully established to infect and analyze large numbers of zebrafish embryos providing a new powerful tool for compound testing and drug discovery using a whole animal vertebrate organism.

 JoVE Biology

Rapid Identification of Chemical Genetic Interactions in Saccharomyces cerevisiae

1Department of Biochemistry and Microbiology, University of Victoria

JoVE 52345

Here we present a cost-effective method for defining chemical-genetic interactions in budding yeast. The approach is built on fundamental techniques in yeast molecular biology and is well suited for the mechanistic interrogation of small to medium collections of chemicals and other media environments.

 JoVE Neuroscience

Design, Surface Treatment, Cellular Plating, and Culturing of Modular Neuronal Networks Composed of Functionally Inter-connected Circuits

1Department of Neurobiology, George S. Wise Faculty of Life Sciences, Tel-Aviv University, 2Department of Neuroscience and Brain Technologies, Istituto Italiano di Tecnologia, 3School of Electrical Engineering, Tel-Aviv University, 4School of Physics and Astronomy, Tel-Aviv University, 5Department of Informatics, Bioengineering, Robotics and System Engineering, University of Genova

JoVE 52572

This manuscript describes a protocol to grow in vitro modular networks consisting of spatially confined, functionally inter-connected neuronal circuits. A polymeric mask is used to pattern a protein layer to promote cellular adhesion over the culturing substrate. Plated neurons grow on coated areas establishing spontaneous connections and exhibiting electrophysiological activity.

 JoVE Environment

Automated Gel Size Selection to Improve the Quality of Next-generation Sequencing Libraries Prepared from Environmental Water Samples

1Department of Pathology and Laboratory Medicine, Faculty of Medicine, The University of British Columbia, 2Coastal Genomics, 3British Columbia Public Health Microbiology and Reference Laboratory

JoVE 52685

This manuscript describes an automated gel size selection approach for purifying DNA fragments for next-generation sequencing. The Ranger Technology provides complete automation of the entire process of agarose gel loading, electrophoretic analysis, and recovery of targeted DNA fragments allowing for high-throughput and high quality next-generation sequencing libraries.

 JoVE Behavior

Early Metamorphic Insertion Technology for Insect Flight Behavior Monitoring

1Department of Electrical and Computer Engineering, North Carolina State University

JoVE 50901

We present a novel surgical procedure to implant electrodes in Manduca sexta during its early metamorphic stages. This technique allows mechanically stable and electrically reliable coupling with the neuromuscular tissue to study flight neurophysiology dynamics. We also present a novel magnetic levitation platform for tethered studies of insect yaw.

 JoVE Biology

Competitive Genomic Screens of Barcoded Yeast Libraries

1Banting and Best Department of Medical Research and Department of Molecular Genetics, University of Toronto, 2Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, 3Donnelly Sequencing Centre, University of Toronto, 4Genetics and Molecular Biology Branch, National Human Genome Research Institute, NIH, 5Stanford Genome Technology Center, Stanford School of Medicine, Stanford University, 6Department of Pharmaceutical Sciences, University of Toronto

JoVE 2864

We have developed comprehensive, unbiased genome-wide screens to understand gene-drug and gene-environment interactions. Methods for screening these mutant collections are presented.

 JoVE Biology

Procedures for Rat in situ Skeletal Muscle Contractile Properties

1Faculty of Kinesiology, University of Calgary

JoVE 3167

This video demonstrates the surgical preparation and procedures needed to study the contractile responses of the rat medial gastrocnemius muscle preparation in situ. This preparation allows measurement of skeletal muscle contractile properties under physiological conditions. The animal is anesthetized and the muscle is separated from surrounding tissue at its distal end. The Achilles tendon is attached to a force transducer, allowing measurement of the muscle’s contractile response at 37 degrees C with an intact circulation.

 JoVE Medicine

High Content Screening in Neurodegenerative Diseases

1Department of Clinical Genetics, VU University Medical Center, 2Center for Neurogenomics and Cognitive Research, Neuroscience Campus Amsterdam

JoVE 3452

We describe a methodology combining automated cell culturing with high-content imaging to visualize and quantify multiple cellular processes and structures, in a high-throughput manner. Such methods can aid in the further functional annotation of genomes as well as identify disease gene networks and potential drug targets.

 JoVE Bioengineering

Planar and Three-Dimensional Printing of Conductive Inks

1Department of Materials Science and Engineering, University of Illinois at Urbana-Champaign, 2Center for Micro- and Nanotechnology, Lawrence Livermore National Laboratory, 3Presently at the Interdisciplinary Center for Wide Band-gap Semiconductors, University Of California Santa Barbara

JoVE 3189

Planar and three-dimensional printing of conductive metallic inks is described. Our approach provides new avenues for fabricating printed electronic, optoelectronic, and biomedical devices in unusual layouts at the microscale.

 JoVE Neuroscience

Muscle Receptor Organs in the Crayfish Abdomen: A Student Laboratory Exercise in Proprioception

1Department of Biology, University of Kentucky

JoVE 2323

The primary purpose of this experiment is to understand how primary sensory neurons convey information of joint movements and positions as proprioceptive information for an animal. An additional objective of this report is present the anatomy of the preparation by dissection and viewing of neurons under a dissecting microscope.

 JoVE Neuroscience

Measurement of Total Calcium in Neurons by Electron Probe X-ray Microanalysis

1Laboratory of Neurobiology, National Institute of Neurological Disorders and Stroke, National Institutes of Health

JoVE 50807

This paper describes the application of cryoanalytical electron microscopy to the quantitative measurement of total calcium content and distribution at subcellular resolution in physiologically defined biological specimens.

 JoVE Biology

High-throughput Purification of Affinity-tagged Recombinant Proteins

1Department of Life Sciences, Imperial College London

JoVE 4110

We describe a method for the affinity-tagged purification of recombinant proteins using liquid-handling robotics. This method is generally applicable to the small-scale purification of soluble His-tagged proteins in a high-throughput format.

 JoVE Engineering

Bringing the Visible Universe into Focus with Robo-AO

1Caltech Optical Observatories, California Institute of Technology, 2Department of Astronomy, California Institute of Technology, 3Dunlap Institute for Astronomy and Astrophysics, University of Toronto, 4Inter-University Centre for Astronomy & Astrophysics, 5Observatories of the Carnegie Institution for Science, 6Benoziyo Center for Astrophysics, Weizmann Institute of Science

JoVE 50021

Light from astronomical objects must travel through the earth's turbulent atmosphere before it can be imaged by ground-based telescopes. To enable direct imaging at maximum theoretical angular resolution, advanced techniques such as those employed by the Robo-AO adaptive-optics system must be used.

 JoVE Neuroscience

Designing and Implementing Nervous System Simulations on LEGO Robots

1Marine Science Center, Northeastern University, 2Fachrichtung Bionik, Bremen University of Applied Sciences

JoVE 50519

An approach to neural network modeling on the LEGO Mindstorms robotics platform is presented. The method provides a simulation tool for invertebrate neuroscience research in both the research lab and the classroom. This technique enables the investigation of biomimetic robot control principles.

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