Wet autoclave method

Tissues are fixed with formalin for 18 hr to 4 weeks and then embedded in paraffin. Sections (4 pm thick) are mounted onto superfrost or poly-L-lysine-coated glass slides, dried in an oven for 1 hr at 60°C, and deparaffmized with three changes of xylene. This is followed by rehydration through a series of descending concentrations of ethanol. The slides are placed 

The following hydrated autoclave method can be employed for immunohistochemical detection of molecules in both cultured cell and tissue specimens. The method was used, for example, to localize androgen receptor in cultured LNCaP cells (derived from prostatic carcinoma metastasized to lymph node) and biopsy specimens from patients with prostatic carcinoma (Ehara et al 1996). After being removed from the culture medium, the cells on plastic cover slips are fixed with 10% formalin for 10 min at 20°C. Tissue specimens are fixed for 1-2 days and embedded in paraffin. Sections (5 pan) are cut, mounted on glass slides, and heated in an oven for 1 hr at 42°C to promote adherence to the slide. After deparaffmizing and rehydration, the sections are subjected to epitope retrieval treatment as follows. 

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