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The Journal of Visualized Experiments (JoVE) is a peer reviewed, PubMed-indexed video journal. Our mission is to increase the productivity of scientific research.

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 JoVE General

Proper Care and Cleaning of the Microscope


JoVE 842 8/11/2008

Core Optical Imaging Facility, University of Texas Health Science Center at San Antonio (UTHSCSA)

Keeping the microscope optics clean is important for high-quality imaging. Dust, fingerprints, excess immersion oil, or mounting medium on or in a microscope causes reduction in contrast and resolution. DIC is especially sensitive to contamination and scratches on the lens surfaces. This protocol details the procedure for keeping the microscope clean.

 JoVE General

Major Components of the Light Microscope


JoVE 843 7/30/2008

Core Optical Imaging Facility, University of Texas Health Science Center at San Antonio (UTHSCSA)

The light microscope is a basic tool for the cell biologist, who should have a thorough understanding of how it works, how it should be aligned for different applications, and how it should be maintained as required to obtain maximum image-forming capacity and resolution. The components of the microscope are described in detail here.

 JoVE General

Screening for Amyloid Aggregation by Semi-Denaturing Detergent-Agarose Gel Electrophoresis


JoVE 838 7/16/2008

1,2,3, 1,2,3

1Whitehead Institute for Biomedical Research, 2Department of Biology, MIT - Massachusetts Institute of Technology, 3Howard Hughes Medical Institute

SDD-AGE is a useful technique for the detection and characterization of amyloid-like polymers in cells. Here we demonstrate an adaptation that makes this technique amenable to large-scale applications.

 JoVE General

Making Patch-pipettes and Sharp Electrodes with a Programmable Puller


JoVE 939 10/08/2008

1, 2, 2

1Department of Molecular and Cellular Physiology, Stanford University, 2Department of Molecular and Cellular Physiology, Stanford University School of Medicine

This video shows how to use a programmable puller to make patch pipettes and sharp electrodes for electrophysiology. The same procedure can be used to make a variety of glass tools, including injection needles.

 JoVE General

Pressure-polishing Pipettes for Improved Patch-clamp Recording


JoVE 964 10/22/2008

, ,

Department of Molecular and Cellular Physiology, Stanford University School of Medicine

This is a guide to modifying the shape of glass micropipettes. Specifically, by using heat and air pressure the taper is widened without increasing the tip opening, leading to lower pipette resistance. This is critical to obtain low noise recordings of small cells but is useful in many applications.

 JoVE General

Cellular Toxicity of Nanogenomedicine in MCF-7 Cell Line: MTT assay


JoVE 1191 4/03/2009

1,2, 1,3, 1, 2, 1,3

1Research Center for Pharmaceutical Nanotechnology, Faculty of Pharmacy, Tabriz University (Medical Sciences), 2Gifted and Talented Students Office, Educational Development Center, Tabriz University (Medical Sciences), 3School of Advanced Biomedical Sciences, Tabriz University (Medical Sciences)

The MTT assay is an easy and reproducible colorimetric assay for evaluation of cell viability based on reduction of yellow MTT and production of water insoluble purple formazan. Here, the viability of MCF-7 cells upon treatment of nanogenomedicine has been evaluated.

 JoVE General

A Lectin HPLC Method to Enrich Selectively-glycosylated Peptides from Complex Biological Samples


JoVE 1398 10/01/2009

1, 2, 1, 1, 1, 2, 1, 1, 1, 3, 2, 1, 1

1Obstetrics, Gynecology and Reproductive Sciences, University of California, San Francisco - UCSF, 2Buck Institute for Age Research, 3Department of Chemistry, Purdue University

Lectin-conjugated POROS beads were employed for HPLC. Glycopeptide standards served as positive and negative controls. MARS-14 depleted, trypsin-digested human plasma was chromatographed and flow-through (FT) and bound fractions collected for ESI-LC-MS/MS analyses. Glycopeptides were enriched in the bound fraction as compared to FT.

 JoVE General

Assembly, Loading, and Alignment of an Analytical Ultracentrifuge Sample Cell


JoVE 1530 11/05/2009

, , ,

National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, Dynamics of Macromolecular Assembly, Laboratory of Bioengineering and Physical Science

The analytical ultracentrifuge (AUC) sample cell holds sample and reference buffer and during experiments and is exposed to high vacuum and rotor speeds up to 60,000 rpm. This video will demonstrate the rigorous attention to detail necessary for assembly, loading and alignment of this very important component of an AUC experiment.

 JoVE General

In vivo Imaging of Intact Drosophila Larvae at Sub-cellular Resolution


JoVE 2249 9/10/2010

*1,2, *1, 1,2, 1, 1, 1

1Junior Research Group Synaptic Plasticity, Hertie Institute for Clinical Brain Research, University of Tübingen, 2Graduate School of Cellular and Molecular Neuroscience, University of Tübingen

This protocol describes a reliable method for anesthetization and imaging of intact Drosophila melanogaster larvae. We have utilized the volatile anesthetic desflurane to allow for repetitive imaging at sub-cellular resolution and re-identification of structures for up to a few days1.

 JoVE General

Modeling Biological Membranes with Circuit Boards and Measuring Electrical Signals in Axons: Student Laboratory Exercises


JoVE 2325 1/18/2011

1, 1, 2, 1

1Department of Biology, University of Kentucky, 2Department of Physiology, University of Toronto

This is a demonstration of how biological membranes can be understood using electrical models. We also demonstrate procedures for recording action potentials from the ventral nerve cord of the crayfish for student orientated laboratories.

 JoVE General

Paraffin-Embedded and Frozen Sections of Drosophila Adult Muscles


JoVE 2438 12/27/2010

, , , , ,

Gene Expression and Signaling Research Group, Max Planck Institute for Biophysical Chemistry

Identification of mechanisms underlying muscle damage is crucial. Here we present the histological technique for preparing paraffin-embedded and frozen sections of Drosophila thoracic muscles. This allows analysis of muscle morphology and localization of protein and other muscle cell components.

 JoVE General

Aseptic Laboratory Techniques: Volume Transfers with Serological Pipettes and Micropipettors


JoVE 2754 5/31/2012

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

When working in a laboratory, it is imperative to minimize sources of contamination. Aseptic technique refers to procedures that permit transfer of cultures and reagents while avoiding contact with non-sterile surfaces. Serological pipettes and micropipettors are used to measure precise volumes without compromising sterility of solutions used in experiments.

 JoVE General

Manual Restraint and Common Compound Administration Routes in Mice and Rats


JoVE 2771 9/26/2012

1, 2, 1, 1, 2

1Insourcing Solutions, Charles River, 2Research Models and Services, Charles River

Working safely and humanely with research rodents requires a core competency in handling and restraint methods. This article will present the basic principles required to safely handle and effectively administer compounds to mice and rats.

 JoVE General

Scale-Up of Mammalian Cell Culture using a New Multilayered Flask


JoVE 3418 12/05/2011

, , , , ,

BD Biosciences

Cells play an instrumental and increasing role in research, and the discovery and development of new therapeutics. With this increasing need for greater number of cells we need more efficient and effective ways for growing and harvesting attachment dependent cells. A Multilayered flask with the right features can serve this purpose.

 JoVE General

Pouring and Running a Protein Gel by reusing Commercial Cassettes


JoVE 3465 2/12/2012

1, 1, 1, 1,2,3

1Department of Biology, University of Florida, 2UF Genetics Institute, University of Florida, 3Plant Molecular & Cellular Biology Program, University of Florida

Our protocol demonstrates how to pour multiple protein gels at a time by recycling Invitrogen Nupage Novex minigel cassettes, and inexpensive materials purchased at a home improvement store. This economical and streamlined method includes a way to store the gels at 4°C for a few weeks. By re-using the plastic gel cassettes from commercially available gels, labs that run frequent protein gels can save significant costs and help the environment.

 JoVE General

Mouse Sperm Cryopreservation and Recovery using the I·Cryo Kit


JoVE 3713 12/12/2011

1, 1, 1, 2

1Genetically Engineered Models and Services, Charles River, 2Research Models and Services, Charles River

Here we demonstrate the newly developed I•Cryo kit for mouse sperm cryopreservation. Two-cell stage embryo development with frozen-thawed sperm was improved consistently in 5 mouse strains with the use of this kit. Over a 1.5 year period, 49 genetically modified mouse lines were archived by sperm cryopreservation with the I•Cryo kit and later successfully recovered by IVF.

 JoVE General

Multiplexed Fluorometric ImmunoAssay Testing Methodology and Troubleshooting


JoVE 3715 12/12/2011

, , ,

Research Animal Diagnostic Services (RADS), Charles River

Using Luminex Corporation’s xMAP microsphere technology, we have developed the Multiplexed Fluorometric ImmunoAssay (MFIA) for serosurveillance of various laboratory animal species. The MFIA is a suspension microarray where antigen, tissue control or immunoglobulins are covalently linked to color-coded polystyrene microspheres. The MFIA testing method as well as various troubleshooting topics is addressed.

 JoVE General

Do-It-Yourself Device for Recovery of Cryopreserved Samples Accidentally Dropped into Cryogenic Storage Tanks


JoVE 3903 5/11/2012

1,2, 1,2,3, 1,2

1Molecular and Microbiology Department and Center for the Study of Genomics in Liver Diseases, George Mason University, 2Translational Research Institute, Inova Health System, 3Research Center for Medical Genetics RAMS

Here we present a low cost, durable cryotolerant device for sample retrieval from Dewar tanks filled with liquid nitrogen. The ease of construction and modular design of the device makes the process of sample retrieval from cryogenic tanks safe and easy.

 JoVE General

Multi-parameter Measurement of the Permeability Transition Pore Opening in Isolated Mouse Heart Mitochondria


JoVE 4131 9/07/2012

, , ,

Department of Anesthesiology & Pain Medicine, Mitochondria and Metabolism Center, University of Washington, Seattle

A spectrofluorometric protocol for the measurement of the mitochondrial permeability transition pore opening in isolated mouse heart mitochondria is presented here. The assay involves the simultaneous measurement of mitochondria Ca2+ handling, mitochondrial membrane potential and mitochondrial volume. The procedure for obtaining high-quality and functional heart mitochondria is also described.

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