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Reflections on the use of controls in immunohistochemistry and proposal for application of a multitissue spring-roll
Insights
This study introduces a cost-effective multitissue control block for immunohistochemistry, improving accuracy by running controls on the same slide as test tissues. This method enhances reliability and reduces diagnostic errors in pathology labs.
Area of Science:
- Pathology
- Immunohistochemistry
- Diagnostic Laboratory Science
Background:
- Conventional immunohistochemistry (IHC) positive controls are separate slides, which cannot confirm correct reagent application on test cases.
- Current positive controls are expensive, comprising 20-40% of slides in diagnostic surgical pathology labs.
- The efficacy of conventional negative controls in IHC is also questioned.
Purpose of the Study:
- To describe a novel, cost-effective method for creating reliable immunohistochemistry controls.
- To enhance the assurance of proper immunostaining procedures, sensitivity, and specificity on individual slides.
Main Methods:
- Development of a simple method to create a multitissue spring-roll control block using mostly normal tissues.
- Mounting multitissue controls on the same slides as the test cases.
- Subjecting both control and test tissues to identical immunostaining procedures.
Main Results:
- The multitissue controls, processed identically to test tissues, provide superior assurance of immunostain performance.
- This integrated approach verifies correct reagent use and staining parameters for each individual slide.
- The method offers a more reliable assessment of sensitivity and specificity compared to conventional controls.
Conclusions:
- The multitissue spring-roll control block is a superior and more economical alternative to conventional positive controls in IHC.
- This method ensures the reliability and accuracy of immunohistochemical staining in diagnostic pathology.
- The presence of multiple immunostains on a single case can adequately indicate nonspecific staining, questioning the need for separate negative controls.
Abstract:
The conventional positive control in immunohistochemistry consists of a separately immunostained slide containing tissue known to show positive reaction with the antibody. Although it can indicate that the reagents are sound, it cannot guarantee that the appropriate reagents have indeed been dispensed in the correct order on the test cases. Furthermore, positive controls are costly because they often account for 20% to 40% of all immunostained slides in a diagnostic surgical pathology laboratory. We describe a simple method to produce a multitissue spring-roll control block, comprising mostly normal tissues. The multitissue controls are mounted on the same slides of the test cases, and thus undergo the identical immunostaining procedures as the test tissues. Therefore they can provide the best assurance that the immunostains are working properly on the individual slides, the expected sensitivity is achieved, and the specificity is as expected. The wisdom of the conventional negative control is also questioned. When two or more immunostains are performed on a case, they should be sufficient to indicate the presence or absence of nonspecific staining.
