Fig. 1

Comparison of two mIHC protocols for use in human FFPE tissue. (A) The multiple interactive labeling by antibody neodeposition (MILAN) method uses β-mercaptoethanol and sodium dodecyl sulfate (βME + SDS) to strip the antibody complex. (B) The multiplexed immunohistochemical consecutive staining on a single slide (MICSSS) uses ethanol (EtOH) to wash out the chromogen. Heat-induced epitope retrieval (HIER) is predicted to elute the antibody partially. (C, D) GFAP and IBA1 were stained on serial sections of human FFPE brain tissue. Following the MILAN or MICSSS procedure the slide were re-imaged. (E) 20% of GFAP staining and 90% of IBA1 staining were found across the entire tissue section following the MILAN method. (F) The MICSSS method effectively reduced the re-development of GFAP and IBA1 to less than 0.1%