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Kidney, Artificial: Device(s) which can substitute for normally functioning kidneys in cleansing the blood.
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Using Bioluminescent Imaging to Investigate Synergism Between Streptococcus pneumoniae and Influenza A Virus in Infant Mice

1Department of Microbiology and Immunology, University of Melbourne, 2Laboratory of Pediatric Infectious Diseases, Radboud University Nijmegen Medical Centre, 3The Centre for Dynamic Imaging, The Walter and Eliza Hall Institute for Medical Research

JoVE 2357


 Immunology and Infection

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A Simple Composite Phenotype Scoring System for Evaluating Mouse Models of Cerebellar Ataxia

1Department of Biochemistry, University of Washington, 2Department of Neurology, University of Washington, 3Division of Genetics, Departments of Pediatrics and Cellular and Molecular Medicine, and the Institute for Genomic Medicine, University of California, San Diego - Rady Children’s Hospital

JoVE 1787


 Neuroscience

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Making Solutions in the Laboratory

JoVE 5030

The ability to successfully make solutions is a basic laboratory skill performed in virtually all biological and chemical experiments. A solution is a homogenous mixture of solute dissolved in bulk liquid known as the solvent. Solutions can be described by their solute concentration, a measure of how much solute is present per unit of solution. In this video, a step-by-step procedure for how to make a water-based, or aqueous, solution for biological applications is presented. The video discusses how to calculate and measure the amount of solute needed for a given volume of solution. Methods for dissolving the solute in purified water and adjusting the pH of the solution are shown. Proper addition of the quantity sufficient (QS) to reach the desired volume is demonstrated with respect to the meniscus before discussing methods for sterilizing the solution. Applications of making solutions are presented through the discussion of several commonly used biological solutions, such as phosphate buffered saline (PBS), and their uses in biological research. These solutions are buffers that mimic physiological pH and osmolarity of cellular fluids.


 General Laboratory Techniques

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Genetic Screens

JoVE 5542

Genetic screens are critical tools for defining gene function and understanding gene interactions. Screens typically involve mutating genes and then assessing the affected organisms for phenotypes of interest. The process can be “forward”, where mutations are generated randomly to identify unknown genes responsible for the phenotypes, or it can be “reverse”, where specific genes are targeted for mutation to observe what phenotypes are produced.Here, JoVE reviews various types of genetic screens, including those that depend on either loss-of-function or gain-of-function mutations, which respectively decrease or increase the activity of genes. We then explore general protocols for forward and reverse screens in a popular model organism, the nematode worm. Finally, we highlight how screens are applied in research today, for example to better understand gene interactions that may contribute to neurodegenerative diseases.


 Genetics

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Annexin V and Propidium Iodide Labeling

JoVE 5650

Staining with annexin V and propidium iodide (PI) provides researchers with a way to identify different types of cell death—either necrosis or apoptosis. This technique relies on two components. The first, annexin V, is a protein that binds certain phospholipids called phosphatidylserines, which normally occur only in the inner, cytoplasm-facing leaflet of a cell’s membrane, but become “flipped” to the outer leaflet during the early stages of apoptosis. The second component is the DNA-binding dye molecule PI, which can only enter cells when their membranes are ruptured—a characteristic of both necrosis and late apoptosis.This video article begins with a review of the concepts behind annexin V and PI staining, and emphasizes how differential patterns of staining can be used to distinguish between cells progressing down different death pathways. We then review a generalized protocol for this technique, followed by a description of how researchers are currently using annexin V and PI staining to better understand cell death.


 Cell Biology

12526
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