Variables Affecting IHC

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Variables Affecting IHC

All factors related to each testing phase of specimen processing in the pathology laboratory may affect IHC staining.
Critical variables that affect staining results include:
  • Temperature
  • pH
  • Ionic conditions
  • Titer of antibody concentration
  • Number of epitope sites per cell
  • Duration of incubation
  • Fixation and tissue processing
Quality Control (QC) Factors by Test Phase (not a complete list)
Pre AnalyticAnalyticPost Analytic
Fixation: Time, type, volumeHeating, dewax, pretreatmentsLocalization, interpretation
Tissue: Type, dimensionAntigen retrievalPositive-negative tissue controls: Cut off values or scoring
Tissue processing completePrimary AB, dilution, clone, time, temperatureReport elements (eg, synoptic)
Decalcification: Type, timeDetection: Sensitivity, selectivity FDA disclaimers
Section thickness, drying, storageControls