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Published on: September 10, 2017
Effect of storage on p63 immunohistochemistry: a time-course study
Holly N Burford1, Amy L Adams, Omar Hameed
1Department of Pathology, University of Alabama School of Medicine, Birmingham, AL 35294-6823, USA.
Insights
Storage of unstained slides negatively impacts p63 immunohistochemistry staining intensity. This time-dependent decline, evident within two weeks, affects diagnostic accuracy in surgical pathology and research.
Area of Science:
- Pathology
- Immunohistochemistry
- Oncology
Background:
- Immunohistochemistry (IHC) staining intensity can be affected by unstained slide storage.
- The impact of storage on p63 antigen staining intensity is not well-documented.
Purpose of the Study:
- To evaluate the effect of storage duration on p63 immunohistochemical staining intensity.
- To determine the time course of staining decline in unstained p63 slides.
Main Methods:
- A time-course study was conducted on breast lesion slides.
- Slides were stained for p63 at baseline and after 2, 4, and 8 weeks of room temperature storage.
- A p63 intensity score was calculated based on staining intensity and percentage of positive myoepithelial cells.
Main Results:
- A progressive decline in p63 staining intensity score was observed over time.
- Statistically significant differences in staining intensity were found between day 0 and 2 weeks (P=0.037), and 2 and 4 weeks (P=0.001).
- The decline between 4 and 8 weeks was not statistically significant (P=0.075).
Conclusions:
- Unstained slide storage leads to a time-dependent decrease in p63 immunostaining intensity.
- This decline is detectable as early as 2 weeks and persists up to 8 weeks.
- Consideration of storage time is crucial for accurate interpretation of p63 immunostains in diagnostics and research.
Abstract:
One of the problems with immunohistochemistry is that storage of unstained slides can have a detectable negative effect on the intensity of staining for various antigens. This has not been well studied for the p63 antigen. A time-course study was performed to evaluate the effect of storage on the intensity of p63 immunohistochemical staining. Slides from various breast lesions/foci were stained with a p63 antibody on day 0 and after storage at room temperature for 2, 4, and 8 weeks. A p63 intensity score, ranging from 0 to 300, was calculated using the intensity (0, 1, 2, or 3+) and percentage of peripheral myoepithelial cell staining (0% to 100%). There was a progressive decline in the p63 intensity score with time with the differences between day 0 and 2 weeks and 2 and 4 weeks being statistically significant (P=0.037 and 0.001, respectively), whereas that between 4 and 8 weeks was not (P=0.075). These findings confirm a time-dependent decline in p63 immunostaining intensity evident as early as with 2 weeks of storage and still present after 8 weeks. This should be considered in the evaluation of p63 immunostains performed on stored unstained sections, both in diagnostic surgical pathology and in translational research studies.
