Electron Microscopy

The resolution of a microscope is limited by the wavelength of light passing through the sample. For visible microscopes using 400 nm light (blue light), the limit of resolution is one half the wavelength, or 200nm. This is some two to three orders of magnitude larger than many cellular structures. Electrons, like photons, have wavelike…

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Detection Systems in Immunohistochemistry

Light microscopy makes use of primarily two detection systems for immunohistochemistry – fluorescence and enzyme labeling, while electron microscopy relies on the deposition of electron dense materials at the site of antibody binding. Techniques for light microscopy are discussed below. EM is covered briefly in the next section. Immunoflourescence The conjugation of a fluorescent dye…

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Antibody Binding

The antibody systems used in Immunohistochemistry can be broadly divided into two types, direct and indirect. With the direct method the visualizing agent is attached directly to the antibody that will bind with the antigen. The direct method is technically and theoretically straightforward, and yields results sufficient for many studies. Its sensitivity is limited by…

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Staining Procedures

Most dyes used to visualize the membranes and organelles of the cell are water soluble. The embedded wax must therefore be removed prior to staining. This is done by effectively reversing the tissue processing schedule. There are literally thousands of staining protocols and procedures in use. As an example, one of the most common stains,…

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