Histology & Immunohistochemistry Protocol Development & Validation

Protocols for histology and immunohistochemistry are established in the lab following standard laboratory practices. For new targets, appropriate positive and negative controls are identified. These controls are used to determine the optimal experimental conditions. For example, appropriate IHC and IF conditions are determined by staining a collection archival tissue and cell pellets that are known to either strongly express or not express a specific antigen. This is often achieved by constructing tissue microarrays, which reduces experiment-to-experiment variability. Doing this defines the range of IHC levels for a given antigen or antibody target.

All protocol development is done using automated equipment (e.g., DAKO Autostainer, etc) to further reduce run-to-run variability. IHC and IF stains are reviewed by a board certified pathologist to ensure high levels of quality assurance (QA) and quality control (QC). Once suitable histology conditions are established and validated, the corresponding protocol is written and reviewed by the CTRL lab director at which point it becomes the standard operating procedure.

Any subsequent modifications are initially validated on appropriate TMA(s) and previously stained samples before the protocol is formally revised.