Complete one-hour immunocytochemistry based on capillary action

We describe a less than one-hour manual method for immunocytochemical analyses of B5 or formalin-fixed, paraffin-embedded tissue sections. The method employs capillary action to sequentially apply, incubate and remove liquid reagents from apposed pairs of up to 20 glass microscope slides and allows for simultaneous immunocytochemical analyses of as many as 10 different antigens. The method described here uses a) positively charged glass slides to rapidly immobilize tissue sections; b) rapid deparaffinization techniques; c) multipurpose reagents; d) ethanol-enriched buffer washes to improve capillary action and reduce nonspecific background; e) a single broad spectrum streptavidin-peroxidase or streptavidin-alkaline phosphatase detection system that identifies most primary monoclonal and polyclonal antibodies; and f) specific immunocytochemical signal amplification by cyclic chromogen enhancement.