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Immunohistochemistry and Special Stain Requisition 315 Erin Drive, Knoxville TN 37919 865.584.1933 800.772.0951 fax 865.584.1323 www.kdlpathology.com Knoxville Dermatopathology Laboratory Dedicated

How to fill out immunohistochemistry and special stain

How to fill out immunohistochemistry and special stain:

1. Gather all necessary materials and reagents required for the selected immunohistochemistry or special stain protocol. This may include tissue samples, antibodies, detection reagents, staining solutions, and control slides.
2. Prepare the tissue sections for staining by ensuring they are properly fixed, embedded, and sectioned. Follow the appropriate protocols for the specific type of tissue being stained.
3. Perform any necessary antigen retrieval steps to expose the target antigens for proper antibody binding. This may involve heating, enzymatic digestion, or other techniques depending on the specific antigen and staining protocol.
4. Block non-specific antibody binding sites by incubating the tissue sections with a blocking solution. This prevents false positive staining and improves specificity.
5. Apply the primary antibody to the tissue sections and incubate for the recommended time and temperature. The primary antibody should specifically target the antigen of interest.
6. Wash the tissue sections thoroughly to remove any unbound primary antibody.
7. Apply the secondary antibody conjugated to a detection system, such as a fluorescent dye or enzyme, that will allow visualization of the primary antibody. Incubate for the recommended time and temperature.
8. Wash the tissue sections again to remove any unbound secondary antibody.
9. If using an enzyme detection system, apply the appropriate substrate solution that will generate a visible signal. Incubate for the specified time and monitor for color development.
10. Wash the tissue sections one final time to remove any excess substrate or reagents.
11. Mount the stained sections on glass slides using a mounting medium that preserves the tissue and allows for microscopic examination.
12. Cover the slides with coverslips and seal with an appropriate mounting medium to protect the stained sections and prevent fading.
13. Analyze the stained sections using a microscope equipped with the appropriate filters or imaging system. Capture and document the images if necessary.
14. Interpret the staining results based on the specific staining protocol, control slides, and any relevant literature or guidelines.

Who needs immunohistochemistry and special stain?

1. Immunohistochemistry and special stains are often used in medical and research fields, particularly in pathology and histology.
2. Pathologists use immunohistochemistry and special stains to aid in the diagnosis of diseases and to differentiate between various tissue types and structures.
3. Researchers use these techniques to investigate the expression and localization of specific proteins, antigens, or molecules within tissues, helping to elucidate their roles in both normal and disease states.
4. Immunohistochemistry and special stain techniques are also valuable in pharmaceutical and biotechnology industries for drug development, quality control, and safety assessment purposes.
5. Veterinary pathologists utilize these techniques to identify diseases in animals and to study animal models of human diseases.

Overall, immunohistochemistry and special stains play a crucial role in understanding tissue morphology, disease processes, and identifying potential therapeutic targets, making them essential tools in both clinical and research settings.

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