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scanning electron microscopy, fluorescence microscopy, and confocal microscopy. There are several different methods used in the study of cells: Cell culture is the basic technique of growing cells in a laboratory independent of an organism. Immunostaining, also known as immunohistochemistry, is a specialized histological method used to localize proteins in cells or tissue slices. Unlike regular histology, which uses stains to identify cells, cellular components or protein classes, immunostaining requires the reaction of an antibody directed against the protein of interest within the tissue or cell. Through the use of proper controls and published protocols (need to add reference links here), specificity of the antibody-antigen reaction can be achieved. Once this complex is formed, it is identified via either a "tag" attached directly to the antibody, or added in an additional technical step. Commonly used "tags" include fluorophores or enzymes. In the case of the former, detection of the location of the "immuno-stained" protein occurs via fluorescence microscopy. With an enzymatic tag, such as horse radish peroxidase, a chemical reaction is carried out that results in a dark color in the location of the protein of interest. This darkened pattern is then detected using light microscopy. Computational genomics is used to find patterns in genomic information [2] DNA microarrays identify changes in transcript levels between different experimental conditions. Gene knockdown mutates a selected gene. In situ hybridization shows which cells are expressing a particular RNA transcript. PCR can be used to determine how many copies of a gene are present in a cell. Transfection introduces a new gene into a cell, usually an expression construct Purification of cells and their parts Purification may be performed using the following methods: Cell fractionation Release of cellular organelles by disruption of cells. Separation of different organelles by centrifugation. Flow cytometry Immunoprecipitation Proteins extracted from cell membranes by detergents and salts or other kinds of chemicals. |
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