Immunofluorescence histochemical direct method

Summary

Immunofluorescent tissue (cell) chemical staining method - Direct method, a fluorescent antibody staining method.

Principle

An antibody labeled with fluorescein of known specificity binds directly to the corresponding antigen in the tissue cells, and specific fluorescence is seen under a fluorescence microscope at the site where the antigen is present.

Appliance

It is suitable for the detection of protein antigens with high concentration, such as the examination of kidney and skin antigens. Also commonly used in clinical practice for the detection of bacteria, spirochetes, protozoa and fungi.

Operation method

Check antigen method; check antibody method

Materials and Instruments

[Materials] Specific antigen; specific antibody;

Move

1. Staining After fixation, the sections were stained with fluorescent antibody diluted to the staining potency for 30 minutes at room temperature or 37~C in a wet box.Wash the sections to remove the fluorescent antibody, immerse the sections in PBS at pH 7.2-7.4 and wash with shaking or vibration for 5 minutes each time, then wash with distilled water for 1 minute to remove salt crystals.3. Sealing and observation The sections were sealed with 50% glycerol buffer (0.5 mol/L phosphate buffer, pH 9.0-9.5) or anti-quenching agent, and then placed under a fluorescence microscope for observation.

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