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Blood Gas Analysis: Measurement of oxygen and carbon dioxide in the blood.

Internal Standards

JoVE 10225

Source: Laboratory of Dr. B. Jill Venton - University of Virginia

The goal of many chemical analyses is a quantitative analysis, where the amount of a substance in a sample is determined. In order to accurately calculate the concentration of an unknown from a sample, careful sample preparation is key. Every time a sample is handled or transferred, some of the sample can be lost. There are strategies however, for minimizing sample loss. There are also strategies for coping with sample loss and still making accurate measurements of concentration. To minimize sample loss, the ideal is to minimize the number of sample handling and transfer steps. For example, massing a solid sample directly into a flask that a solution will be made in reduces a transfer step. If it's necessary to transfer from one flask to another and a dilution is being made, then triple rinsing the glassware helps ensure all the sample is transferred. Other strategies are more specific to the sample. For example, samples that adsorb to glass, such as proteins, might better be handled in polypropylene disposable tubes. The tubes are not hydrophilic, so if a small amount of sample is to be pipetted in water, it is best to have already added the water to the tube, so the sample can be pipetted directly into the solve


 Analytical Chemistry

Development of the Chick

JoVE 5155

The chicken embryo (Gallus gallus domesticus) provides an economical and accessible model for developmental biology research. Chicks develop rapidly and are amenable to genetic and physiological manipulations, allowing researchers to investigate developmental pathways down to the cell and molecular levels.

This video review of chick development begins by describing the process of egg fertilization and formation within the chicken reproductive tract. Next, the most commonly used chick staging nomenclature, the Hamburger Hamilton staging series, is introduced. Major events in chick development are then outlined, including the dramatic cellular movements known as gastrulation that form the three major cell layers: The ectoderm, mesoderm, and endoderm. Cells from these layers go on to generate all the tissues within the organism, as well as extraembryonic membranes, which are necessary for the transport of gases, nutrients, and wastes within the eggshell. To conclude the discussion, some exciting techniques will be presented as strategies for studying chick development in greater detail.


 Biology II

Anesthesia Induction and Maintenance

JoVE 10263

Source: Kay Stewart, RVT, RLATG, CMAR; Valerie A. Schroeder, RVT, RLATG. University of Notre Dame, IN

The Guide for the Care and Use of Laboratory Animals ("The Guide") states that pain assessment and alleviation are integral components of the veterinary care of laboratory animals.1 The definition of anesthesia is the loss of feeling or sensation. It is a dynamic event involving changes in anesthetic depth with respect to an animal's metabolism, surgical stimulation, or variations in the external environment.


 Lab Animal Research

High-frequency Ultrasound Imaging of the Abdominal Aorta

JoVE 10397

Authors: Amelia R. Adelsperger, Evan H. Phillips, and Craig J. Goergen, Weldon School of Biomedical Engineering, Purdue University, West Lafayette, Indiana

High-frequency ultrasound systems are used to acquire high resolution images. Here, the use of a state-of-the-art system will be demonstrated to image the morphology and hemodynamics of small pulsatile arteries and veins found in mice and rats. Ultrasound is a relatively inexpensive, portable, and versatile method for the noninvasive assessment of vessels in humans as well as large and small animals. These are several key advantages that ultraound offers compared to other techniques, such as computed tomography (CT), magnetic resonance imaging (MRI), and near-infrared fluorescence tomography (NIRF). CT requires ionizing radiation and MRI can be prohibitively expensive and even impractical in some scenarios. NIRF, on the other hand, is limited by the penetration depth of light required to excite the fluorescent contrast agents. Ultrasound has limitations in terms of imaging depth; however, this may be overcome by sacrificing resolution and using a lower frequency transducer. Abdominal gas and excess body weight can severely diminish image quality. In the first case, the propagation of sound waves is limited, while in the lat


 Biomedical Engineering

Synthesis of an Oxygen-Carrying Cobalt(II) Complex

JoVE 10430

Source: Deepika Das, Tamara M. Powers, Department of Chemistry, Texas A&M University

Bioinorganic chemistry is the field of study that investigates the role that metals play in biology. Approximately half of all proteins contain metals and it is estimated that up to one third of all proteins rely on metal-containing active sites to function. Proteins that feature metals, called metalloproteins, play a vital role in a variety of cell functions that are necessary for life. Metalloproteins have intrigued and inspired synthetic inorganic chemists for decades, and many research groups have dedicated their programs to modeling the chemistry of metal-containing active sites in proteins through the study of coordination compounds. The transport of O2 is a vital process for living organisms. O2-transport metalloproteins are responsible for binding, transporting, and releasing oxygen, which can then be used for life processes such as respiration. The oxygen-carrying cobalt coordination complex, [N,N'-bis(salicylaldehyde)ethylenediimino]cobalt(II) [Co(salen)]2 has been studied extensively to gain understanding about how metal complexes reversibly bind O2.1 In this experiment, we will synthesize [Co


 Inorganic Chemistry

Cardiac Magnetic Resonance Imaging

JoVE 10393

Source: Frederick W. Damen and Craig J. Goergen, Weldon School of Biomedical Engineering, Purdue University, West Lafayette, Indiana

In this video, high field, small-bore magnetic resonance imaging (MRI) with physiological monitoring is demonstrated to acquire gated cine loops of the murine cardiovascular system. This procedure provides a basis for assessing left-ventricular function, visualizing vascular networks, and quantifying motion of organs due to respiration. Comparable small animal cardiovascular imaging modalities include high-frequency ultrasound and micro-computed tomography (CT); however, each modality is associated with trade-offs that should be considered. While ultrasound does provide high spatial and temporal resolution, imaging artifacts are common. For example, dense tissue (i.e., the sternum and ribs) can limit imaging penetration depth, and hyperechoic signal at the interface between gas and liquid (i.e., pleura surrounding the lungs) can blur contrast in nearby tissue. Micro-CT in contrast does not suffer from as many in-plane artifacts, but does have lower temporal resolution and limited soft-tissue contrast. Furthermore, micro-CT uses X-ray radiation and often requires the use of contrast agents to visualize vasculature, both of which are known to cause side effects at high doses incl


 Biomedical Engineering

Introduction to Mass Spectrometry

JoVE 5634

Source: Laboratory of Dr. Khuloud Al-Jamal - King's College London

Mass spectrometry is an analytical chemistry technique that enables the identification of unknown compounds within a sample, the quantification of known materials, the determination of the structure, and chemical properties of different molecules.

A mass spectrometer is composed of an ionization source, an analyzer, and a detector. The process involves the ionization of chemical compounds to generate ions. When using inductively coupled plasma (ICP), samples containing elements of interest are introduced into argon plasma as aerosol droplets. The plasma dries the aerosol, dissociates the molecules, and then removes an electron from the components to be detected by the mass spectrometer. Other ionization methods such as electrospray ionization (ESI) and matrix assisted laser desorption ionization (MALDI) are used to analyze biological samples. Following the ionization procedure, ions are separated in the mass spectrometer according to their mass-to-charge ratio (m/z), and the relative abundance of each ion type is measured. Finally, the detector commonly consists in an electron multiplier where the collision of ions with a charged anode leads to a cascade of increasing number of electrons, which can b


 Analytical Chemistry

Normothermic Ex Situ Heart Perfusion in Working Mode: Assessment of Cardiac Function and Metabolism

1Department of Surgery, Faculty of Medicine, University of Alberta, 2Department of Pediatrics, Faculty of Medicine, University of Alberta, 3Department of Biomedical Engineering, Faculty of Medicine, University of Alberta, 4Department of Mechanical Engineering, Faculty of Engineering, University of Alberta, 5Department of Chemical and Materials Engineering, Faculty of Engineering, University of Alberta, 6Department of Physiology, Faculty of Medicine, University of Alberta, 7Canadian National Transplant Research Program

Video Coming Soon

JoVE 58430


 JoVE In-Press

Diagnostic Necropsy and Tissue Harvest

JoVE 10294

Source: Kay Stewart, RVT, RLATG, CMAR; Valerie A. Schroeder, RVT, RLATG. University of Notre Dame, IN

Many animal experiments rely on final data collection time points that are gathered from the harvesting and testing of organs and tissues. The use of appropriate methods for the collection of organs and tissues can impact the quality of the samples and the analysis of the data that is gleaned for the testing of the tissues. The method of euthanasia of the animal can also impact the quality of the samples. This manuscript will outline proper necropsy techniques for rats.


 Lab Animal Research

Lead Analysis of Soil Using Atomic Absorption Spectroscopy

JoVE 10021

Source: Laboratories of Margaret Workman and Kimberly Frye - Depaul University

Lead occurs naturally in soil, in levels ranging from 10-50 ppm. However, with the widespread use of lead in paint and gasoline in addition to contamination by industry, urban soils often have concentrations of lead significantly greater than background levels – up to 10,000 ppm in some places. Ongoing problems arise from the fact that lead does not biodegrade, and instead remains in the soil. Serious health risks are associated with lead poisoning, where children are particularly at risk. Millions of children in the U.S. are exposed to soil containing lead. This exposure can cause developmental and behavioral problems in children. These problems include learning disabilities, inattention, delayed growth, and brain damage. The Environmental Protection Agency has set a standard for lead in soil at 400 ppm for play areas and 1,200 ppm for non-play areas. Lead is also of concern in soil, when it’s used for gardening. Plants take up lead from the soil. Therefore, vegetables or herbs grown in contaminated soil can lead to lead poisoning. In addition, contaminated soil particles can be breathed in while gardening or brought into the house on clothing and footwear. It is recommended that s


 Environmental Science

Ferric Chloride-induced Canine Carotid Artery Thrombosis: A Large Animal Model of Vascular Injury

1Department of Neurological Surgery, Ohio State University, 2Department of Surgery, Ohio State University, 3Department of Surgery, Duke University, 4Department of Internal Medicine, Ohio State University

JoVE 57981


 Medicine

An All-on-chip Method for Rapid Neutrophil Chemotaxis Analysis Directly from a Drop of Blood

1Institute of Applied Technology, Hefei Institutes of Physical Science, Chinese Academy of Sciences, 2University of Science and Technology of China, 3Department of Physics and Astronomy, University of Manitoba, 4Department of Biosystems Engineering, University of Manitoba, 5Seven Oaks General Hospital, 6Department of Immunology, University of Manitoba, 7Department of Biological Sciences, University of Manitoba

JoVE 55615


 Immunology and Infection

Determining the Solubility Rules of Ionic Compounds

JoVE 10197

Source: Laboratory of Dr. Neal Abrams — SUNY College of Environmental Science and Forestry

An ionic compound's solubility can be determined via qualitative analysis. Qualitative analysis is a branch of analytical chemistry that uses chemical properties and reactions to identify the cation or anion present in a chemical compound. While the chemical reactions rely on known solubility rules, those same rules can be determined by identifying the products that form. Qualitative analysis is not typically done in modern industrial chemistry labs, but it can be used easily in the field without the need of sophisticated instrumentation. Qualitative analysis also focuses on understanding ionic and net ionic reactions as well as organizing data into a flow chart to explain observations and make definitive conclusions. Many cations have similar chemical properties, as do the anion counterparts. Correct identification requires careful separation and analysis to systematically identify the ions present in a solution. It is important to understand acid/base properties, ionic equilibria, redox reactions, and pH properties to identify ions successfully. While there is a qualitative test for virtually every elemental and polyatomic ion, the identification process typically begi


 General Chemistry

Autologous Endothelial Progenitor Cell-Seeding Technology and Biocompatibility Testing For Cardiovascular Devices in Large Animal Model

1Department of Biomedical Engineering, Duke University, 2School of Medicine, Duke University, 3Department of Surgery, Duke University Medical Center, 4School of Medicine, University of Pennsylvania

JoVE 3197


 Bioengineering

Observational Study Protocol for Repeated Clinical Examination and Critical Care Ultrasonography Within the Simple Intensive Care Studies

1Department of Critical Care, University of Groningen, University Medical Centre Groningen, 2Department of Anesthesiology, University of Groningen, University Medical Centre Groningen, 3Department of Radiology, University of Groningen, University Medical Centre Groningen, 4Department of Cardiology, University of Groningen, University Medical Centre Groningen, 5Division of Nephrology, Department of Internal Medicine, University of Groningen, University Medical Centre Groningen

Video Coming Soon

JoVE 58802


 JoVE In-Press

Recording Brain Electromagnetic Activity During the Administration of the Gaseous Anesthetic Agents Xenon and Nitrous Oxide in Healthy Volunteers

1Centre for Human Psychopharmacology, Swinburne University of Technology, 2Department of Anaesthesia and Pain Management, St. Vincent's Hospital Melbourne, 3Brain and Psychological Science Research Centre, Swinburne University of Technology, 4Department of Anaesthesiology, University of Auckland

JoVE 56881


 Neuroscience

Efficiency of Liquid-liquid Extraction

JoVE 10426

Source: Kerry M. Dooley and Michael G. Benton, Department of Chemical Engineering, Louisiana State University, Baton Rouge, LA

Liquid-liquid extraction (LLE) is a separation technique used instead of distillation when either: (a) the relative volatilities of the compounds to be separated are very similar; (b) one or more of the mixture components are temperature sensitive even near ambient conditions; (c) the distillation would require a very low pressure or a very high distillate/feed ratio.1The driving force for mass transfer is the difference in solubility of one material (the solute) in two other immiscible or partially miscible streams (the feed and the solvent). The feed and solvent streams are mixed and then separated, allowing the solute to transfer from the feed to the solvent. Normally, this process is repeated in successive stages using counter-current flow. The solute-rich solvent is called the extract as it leaves, and the solute-depleted feed is the raffinate. When there is a reasonable density difference between the feed and solvent streams, extraction can be accomplished using a vertical column, although in other cases a series of mixing and settling tanks may be used.

Complete and Partial Aortic Occlusion for the Treatment of Hemorrhagic Shock in Swine

1Department of Surgery, University of Michigan, 2Hays Innovations

JoVE 58284


 Medicine

Generation of Integration-free Induced Pluripotent Stem Cells from Human Peripheral Blood Mononuclear Cells Using Episomal Vectors

1State Key Laboratory of Experimental Hematology, Institute of Hematology and Blood Disease Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, 2Division of Regenerative Medicine, Department of Medicine, Loma Linda University, 3Department of Orthopaedic Surgery, Loma Linda University, 4Center for Stem Cell Medicine, Chinese Academy of Medical Sciences, 5Department of Stem Cell & Regenerative Medicine, Peking Union Medical College, 6Collaborative Innovation Center for Cancer Medicine, 7Tianjin Key Laboratory of Blood Cell Therapy and Technology

JoVE 55091


 Developmental Biology

Chronic Implantation of Whole-cortical Electrocorticographic Array in the Common Marmoset

1Laboratory for Molecular Analysis of Higher Brain Function, RIKEN Center for Brain Science, 2Laboratory for Marmoset Neural Architecture, RIKEN Center for Brain Science, 3Department of Physiology, Keio University School of Medicine, 4Department of Ultrastructural Research, National Center of Neurology and Psychiatry

Video Coming Soon

JoVE 58980


 JoVE In-Press

Visualization of Amyloid β Deposits in the Human Brain with Matrix-assisted Laser Desorption/Ionization Imaging Mass Spectrometry

1Department of Life and Medical Systems, Doshisha University, 2Bruker Daltonics K.K., 3The Brain Bank for Aging Research, Tokyo Metropolitan Geriatric Hospital and Institute of Gerontology, 4Graduate School of Brain Science, Doshisha University

Video Coming Soon

JoVE 57645


 JoVE In-Press

In Vitro Method to Control Concentrations of Halogenated Gases in Cultured Alveolar Epithelial Cells

1Department of Perioperative Medicine, CHU Clermont-Ferrand, 2Centre National de la Recherche Scientifique Unité Mixte de Recherche (CNRS UMR) 6293, Institut National de la Santé et de la Recherche Médicale (INSERM) U1103, Laboratoire de Génétique, Reproduction et Développement (GReD), Université Clermont Auvergne, 3Department of Pharmacology, CHU Clermont-Ferrand, 4Nurse Anesthetist School, CHU Clermont-Ferrand

JoVE 58554


 Medicine

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