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Their hearts were too full for them to speak to one another, and they dared say nothing to James. Oh, she s behaved shamefully. There can be no excuse for her. I don t understand, replied James, growing pale.

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There are also physical and chemical indicators that can be used to ensure decontamination effectiveness of autoclaves. Stainless steel loggers are commonly used for monitoring temperature of autoclave. Check out how to use it to maintain the maximum performance of your autoclave machine. There are physical, chemical, and biological indicators that can be used to ensure decontamination effectiveness of autoclaves. Bioburden Test of BSC. Biological Safety Cabinets BSC is an enclosed, ventilated laboratory workspace which provides the most effective primary containment for working with infectious agents.

The BSC is tested for bioburden to ensure the adequate protection.

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A blood smear gives information about the number and shape of blood cells. To perform a blood smear, two very clean slides are required. Drop a small drop of blood on one of the slides. Pull blood forward across slide. Calcium Phosphate Transfection. This method works best in cell lines that are highly transformed and adherent and this technique requires few manipulative steps and maintains high levels of reproducibility from experiment to experiment.

Cationic Polymer Transfection. The cells must be fixed and permeabilized to ensure free access of the antibody to its antigen. Fixation methods fall generally into two classes: organic solvents and cross-linking reagents. This video uses formaldehyde as a cross-linking reagent to fix cells. CellTiter 96R AQueous One Solution Cell Proliferation Assay is a colorimetric method for determining the number of viable cells in proliferation or cytotoxicity assays.

The reagent contains a tetrazolium compound MTS.

The MTS is bioreduced by cells into a colored formazan product which can be measured by the absorbance at nm. By cellular mitochondrial dehydrogenases, WST-1 is bioreduced into formazan which can be measured at an absorbance of nm. Cell Synchronization with Nocodazole at Mitosis. Cell Synchronization is a process by which cells at different stages of the cell cycle in a culture are brought to the same phase.

Nocodazole is one of anticancer drugs that synchronize cells at M-phase. It can interfere with the structure and function of microtubules in interphase and mitotic cells. Chimera RNA Interference. Kumiko Ui-Tei at the University of Tokyo. Clonogenic assay allows one to test the capability of adherent cells to survive and replicate following insult with chemicals or radiation. Count the crystal violet stained colonies which incubated for 9 days with appropriate chemical or radiation dose and calculate the survival rate. The auto gas analyzer is a tool to provide accurate carbon dioxide CO2 measurements in incubators.

The Fyrite gas analyzer is the most widely used instrument for measuring carbon dioxide CO2 levels in incubators. It uses the Orsat method of volumetric analysis involving chemical potassium hydroxide absorption of CO2 gas. Competent Cell Preparation. Competent cells are those that possess more easily altered cell walls that DNA can be passed through easily. Because DNA is hydrophilic, it will not normally pass through membrane. In order to make these cells readily incorporate foreign DNA, they must first be made "competent" to take up foreign DNA.

After washing, unbound antibodies are removed. The more analytes in the sample, the less antibodies will be able to bind to antigens in the well. The signal is then detected using labeled secondary antibodies and the decrease in signal is compared to a control. The major advantage of a competitive ELISA is the ability to use crude or impure samples and still selectively bind any antigen that may be present. Human umbilical vein endothelial cells HUVEC are commonly used as a laboratory model system for the physiological and pharmacological investigations.

Dialysis ultrafiltration is a method to concentrate protein or other macromolecules through a membrane with defined pores. The membranes will have a molecular weight cut-off MWCO. This is the limit of size or range of a protein that can fit through the membrane. DNA sequencing is a technique to determine the order of the nucleotide bases-adenine, guanine, cytosine, and thymine. It is indispensable for basic biological research and discovery. Dot blot is a technique can be used as a semiqualitative method for rapid screening of a large number of samples.

It is for detecting, analyzing, and identifying proteins, similar to the western blot technique but differing in that protein samples are not separated electrophoretically but are spotted through circular templates directly onto the membrane. Electroporation applies a large electric pulse temporarily disturbs the phospholipid bilayer, allowing molecules to pass into the cell. It is usually used in molecular biology as a way of introducing some substance into a cell, such as molecular probes, drugs or pieces of coding DNA.

Emulsification is a process to mix adjuvants and immunogen. Adjuvants are mixed and injected with an antigen to prevent catabolism and help increase the immune response by localizing the antigen for an extended time and attracting the appropriate cells T cells, B cells and APC to interact with it. Ethidium Bromide EtBr is commonly used as a non-radioactive marker for identifying and visualizing nucleic acid bands in electrophoresis.

It is a potent mutagen, so its hazardous properties require special safe handling and disposal procedures. Fluorescent Microscope. A fluorescent microscope is an optical microscope used to study properties of substances using the phenomena of fluorescence and phosphorescence. Specimen can be labeled specifically with a fluorescent molecule and be illuminated with specific wavelength which is absorbed by the fluorophores. Gelatin gel zymography in analyze the MMP is now recognized as the important process in cell migration, invasion or tissue remodeling.

GST Enzymatic Digestion. GST fusion proteins allow convenient affinity purification of many proteins of interest but the GST-tag may become inconvenient in various downstream applications. GST-tagged Protein Purification. Protein purification is a series of processes intended to isolate a single type of protein from a complex mixture. We use GST-fusion protein to purify and detect proteins of interest. The GST-fusion protein can be purified from cells via its high affinity for glutathione. As a mature diagnostic tool, regular blood test is widely adopted to evaluate anemia, leukemia, reaction to inflammation and infections, etc.

The test collects information including the number and types of different types of blood cells, the variation in the size of red blood cells, hematocrit, hemoglobin value, platelet count, mean corpuscular hemoglobin, and the average size of red blood cells. Here we demonstrate a simple operation of hematology test for following clinical diagnosis.

His-tagged Protein Purification. Proteins with histidine tag can be purified and detected easily because the string of histidine residues binds to several types of immobilized metal ions, including nickel, cobalt and copper, under specific buffer conditions. Immunofluorescence is a technique to visualize a specific protein or antigen in cells or tissue sections by binding a specific antibody chemically conjugated with a fluorescent dye.

We use the indirect immunofluorescence staining to perform cells fixed on slides and examine under a fluorescence microscope.

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Immunoprecipitation IP is the technique of precipitating an protein antigen out of solution using an antibody specific to that antigen. The beads are then added to the protein mixture and the proteins that are targeted by the antibodies are captured onto the beads. In the assay, the antigen of interested is immobilized by direct adsorption to the assay plate. Detection of the antigen can then be performed by using a matched set of primary antibody and conjugated secondary antibodies.

Integra CL Cell Cultivation. The CELLine bioreactor is a disposable, two-compartment cultivation device suitable for many cell culture applications. We use CL to produce monoclonal antibodies on a laboratory scale. Cell invasion assay is designed to accelerate the screening process for compounds that influence cell migration through extracellular matrices. Matrigel BD , extracted from the Engelbreth-Holm-Swarm EHS mouse sarcoma, resembles the extracellular matrices is used by cell biologists as a substrate in cell invasion assay. Isotopic labeling is a technique for labeling a substance with a stable or radioactive isotope.

By measuring the radioactivity or the abundance of the isotope, we could made observations of the course through a biologic process. This video shows the procedure for using radioisotope phosphorus to label DNA. Firefly luciferase is widely used as a reporter to study gene expression and other cellular events.

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The Luciferase Assay is an extremely sensitive and rapid reagent for quantitation of firefly luciferase. Through the assay, light is produced by converting the chemical energy of luciferin oxidation and can be measured by a luminometer. Biological materials often are dried to stabilize them for storage or distribution. Lyophilization also called freeze-drying, is one method of drying biological materials that minimizes damage to its internal structure.

Making a Glass Knife. The migration assay also known as the Boyden Chamber Assay is a commonly used test to study the migratory response of endothelial cells. During this assay, cells are placed on the upper layer of a chamber and a solution containing the test agent is placed below the chamber. Following an incubation period, the cells migrated through the membrane are stained and counted.

Mononuclear Cell Isolation from Whole Blood. This procedure to isolate mononuclear cells from whole blood is based on density differences between mononuclear cells and other elements found in the blood sample. Differential migration during centrifugation results in the formation of layers containing different cell types. Mononuclear cells are at the interface between the plasma and the ficoll layer and are recovered by washing steps. Three steps to type the mouse genotype : i isolate the genomic DNA from mouse tails; ii PCR to amplify with specific primers; iii electrophoresis to determine the genotype.

A demonstration on the procedure of using MTT assay to assess the viability and the proliferation of regular cells with absorbance detection at nm is shown. This colorimetric assay measures the reduction of yellow 3- 4,5-dimethythiazolyl -2,5-diphenyl tetrazolium bromide MTT by mitochondrial succinate dehydrogenase. MTT assay allows assessing the viability and the proliferation of cells.

This is a colorimetric assay that measures the reduction of yellow 3- 4,5-dimethythiazolyl -2,5-diphenyl tetrazolium bromide MTT by mitochondrial succinate dehydrogenase. Mycoplasma is highly infectious and cross-contamination commonly occurs when new cells are introduced into laboratories. We use PCR-based method for detection of Mycoplasma weekly. It is a highly sensitive, specific and rapid method to detect mycoplasma contamination in cell cultures. Mycoplasma is a common and serious contamination of cell cultures. Mycoplasma contamination can cause the alteration of the phenotypic cell characteristics.

PCR method allows for fast and reliable identification of mycoplasma contamination in cell cultures. Oil Red O Staining for Adipogenesis. Oil Red O is a lysochrome diazo dye used for demonstrating neutral triglycerides and lipids in tissue. This video shows the procedure of Adipogenesis staining with Oil Red O. Optical Fiber Cleavage. This AbVideo demonstrates the operation of automated fiber cleavage to produce flat and angled cleaves on fibers ranging from 80 microns to 1. Optical fibers are flexible, transparent fibers made of high quality extruded glass silica or plastic.

It is frequently adopted as a waveguide, or "light pipe", to transmit the optical signals. Taking advantages of this kind of property, optical fibers have been used in developing biological sensors in concert with traditional bio-probes such as nucleic acids. Optical Fiber End-Face Analysis. This AbVideo demonstrates the operation of automated interferometric inspection system specifically designed for checking the surface quality and flatness of cleaved, polished or lensed fibers.

Pipette Glass Washing. Pipettes are the laboratory instruments used to transport a measured volume of liquid.

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This video shows you how to wash the reusable glass pipettes. Propidium Iodide Staining. Propidium iodide or PI is an intercalating agent and a fluorescent molecule that can be used to stain DNA. PI is used as a DNA stain for both flow cytometry to evaluate cell viability or DNA content in cell cycle analysis and microscopy to visualize the nucleus and other DNA containing organelles. It can be used to differentiate necrotic, apoptotic and normal cells.

Radioactive Signal Detection. Laser scanning with the FLA, instead of using conventional photography, permits analysis and digital storage. Raw Cell Transmigration. The ability of cell migration is commonly used for the evaluation of cell invasion or wound repair.