Choosing an Immunohistochemistry (IHC) System? A Comparison of 3 IHC Stainers and Polymers from Different Manufacturers.
Qian Dai, Emily S Reisenbichler, Marva S Rasco, Debra K Horton, Omar Hameed. University of Alabama at Birmingham
Background: IHC is routinely used in contemporary pathology practice and its quality depends upon many factors including IHC instruments and detection systems. The aim of this study was to compare the IHC results obtained using different open immunostainers and polymer detection kits.
Design: Different control sections for 9 antigens (CD3, CD20, CD45, CK, CK5/6, KI67, TTF1, BCL1, and S100) were sequentially evaluated by IHC utilizing 3 antibodies (Abs) from different sources [BIOCARE (BC), LABVISION (LV), DAKO (DK)], 3 different immunostainers (BC, LV, DK), and 3 different polymers (BC, LV, DK), generating 243 possible Ab/immunostainer/polymer combinations. The intensity (0-3+) and percentage of cells staining (0-100%) were used to generate an H-score (0-300). Technical issues (loss of tissue, failure of staining, no counterstain) were recorded as absent, minor (still interpretable), or major (not interpretable), while background staining was recorded as present or absent.
Results: There was variation in the staining of the different antigens when utilizing the unique Ab/immunostainer/polymer combinations from the same manufacturer (figure 1), with different overall mean H-scores for BC, LV, and DK (171, 199, and 218, respectively; P=0.019, paired ANOVA).
Comparison of staining between the 3 immunostainers (regardless of Ab/polymer) also showed different overall mean H-scores (175, 199, and 201, respectively; P=0.036, paired ANOVA), as did comparison between the 3 polymers (regardless of Ab/immunostainer) (194, 204 and 177, respectively; P=0.0003, paired ANOVA). The overall incidence of technical problems across immunostainers was 18%, 23%, and 20% for BC, LV, and DK, respectively (P>0.05, X2) with major issues representing 43%, 26% and 19% of these issues, respectively (P>0.05, X2). Background staining was seen in 15%, 14%, and 9% of cases utilizing the BC, LV, and DK polymers (P>0.05, X2).
Conclusions: There are significant differences in staining results obtained using different IHC stainers and polymer detection kits. These findings, as well as others (e.g. cost of reagents, quality and timeliness of technical support), should be taken into consideration when selecting the optimal antibodies/immunostainers/detection systems for IHC.
Monday, February 28, 2011 9:30 AM
Poster Session I Stowell-Orbison/Surgical Pathology/Autopsy Awards Poster Session # 247, Monday Morning