Microbiology Laboratory

DEHYDRATION AND MOUNTING STEPS IN IMMUNOHISTOCHEMISTRY STAINING

Written by MicroDok

Dehydration and mounting is often the last step in the immunohistochemistry staining technique. However, the steps of dehydration and mounting in the immunohistochemistry staining technique are usually followed by microscopy, which allows the researcher or investigator to visualize the stained cells on the slides. In some quarters or textbook, this step of microscopy is often called visualization step. Whichever name it is called, whether microscopy or visualization, the sole aim of microscopy or visualization is to see the antibody-antigen complex or reaction after the whole process of immunohistochemistry staining. Only after microscopy and visualization can the researcher or investigator make an informed choice and conclusion on the integrity and implication of the type of results obtained in the immunohistochemistry staining technique. Fluorescent microscope or confocal microscope can be used for the microscopy. Electron microscopy can also be used in some cases.

In previous sections on immunohistochemistry staining, we looked at:

  • Antibody detection
  • Antibody selection
  • Antibody staining
  • Permeabilization and blocking non specific binding
  • Antigen retrieval protocol
  • Deparaffinization steps

Only the dehydration and mounting steps are highlighted in this section. Please see previous sections of the above techniques on the website for further reading. You can also use the SEARCH BOX on the website for your defined and direct search.

The steps involved in the dehydration and mounting are as follows:

  1. Dehydrate tissue sections by moving the slides through the following solutions at the different times:
  • Wash slides in 2 changes of 80 % alcohol for 1 min at room temperature (RT)
  • Wash slides in 2 changes of 95% alcohol for 1 min at RT
  • Wash slides in 2 changes of 100 % alcohol for 1 min at RT
  • Wash slides in 3 changes of xylene for 1 min at RT
  1. Add mounting media to slides
  2. Apply cover slip to the slide
  3. View under the microscope at recommended magnification

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MicroDok

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