Multiple antigens, retrieval

Figure 5.3 Diagram depicts the further-designed studies to test our hypothesis with respect to standardization of immunohistochemistry based on the antigen retrieval technique exemplified in a multiple direction to draw a conclusion, (a) Periods of formalin fixation, (b) Variable delay of fixation, (c) Storage of FFPE tissue blocks or sections, (d) Variable thickness of FFPE tissue sections, (e) Other variable conditions of processing FFPE tissue blocks. The stereoscopic frame of a cube represents the reliable limitation of quantitative IFIC demonstrated by serial studies as recommended in the text. Reproduced with permission from Shi et al., J. Histochem. Cytochem. 2007 55 105-109.

Automated AR systems have very recendy been introduced, sometimes integrated into or with the automated staining instrument. Antigen retrieval has been in use since the early 1990s. Nevertheless, because of the multiple variables inherent in this technique, standardization has been a challenge. These variables include the size and power of the microwave, the temperature of the retrieval solution, the duration of the retrieval, and the rate of cooling. Other variables include the number of slides and containers in the microwave as well as the size, color, and material of these containers. With this new automation, all these variables are controlled, and this results in more reproducible results. 

Lan HY, Mu W, Nikolic-Paterson DJ, Atkins RC. A novel, simple, reliable, and sensitive method for multiple immunoenzyme staining use of microwave oven heating to block antibody cross reactivity and retrieve antigens. J. Histochem. Cytochem. 1995 43, 97-102. 

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