Three-color IHC Staining On Free-floating Sections From Human Brain

INTRODUCTION

Immunohistochemistry (IHC) on human brain tissue sections is widely used for diagnosis by anatomopathologists and researchers. Indeed this technique allows the visualization of specific antigens and thus their distribution into the cells and their compartments. To handle this type of tissue can be very challenging due to the condition of the brain removal and the quality of the tissue. There is a growing interest on multiplexing IHC and being able to perform this technique on free-floating human brain sections as good as formalin-fixed paraffin-embedded sections on slide could be a step forward to analyze larger and thicker specimen (i.e. 50 μm).

The MULTIVIEW® PLUS (mouse-HRP/rabbit-AP) IHC Kit from Enzo Life Sciences is a non-biotin, one-step detection system suitable for demonstrating the expression of multiple antigens on tissue sections. It enables faster and more consistent staining procedures than traditional two-step methods using biotin and avidin/streptavidin conjugates. Indeed, mouse antibodies are directly tagged with HRP and rabbit antibodies with AP at the same time, all with significantly lower background staining.

The main objective of this study was to validate the use of the MULTIVIEW® PLUS (mouse-HRP/rabbit-AP) IHC Kit and the HIGHDEF ® IHC chromogens to detect multiple staining on human free-floating brain sections. The intensity, sensitivity, and specificity obtained for all staining demonstrate the capacity of the Enzo Life Sciences kit and their chromogen to achieve multiplexing IHC on free-floating tissue sections.

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