Block Staining of Mammalian Tissues with Hematoxylin and Eosin

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Abstract
Various mammalian tissues were stained en bloc with hematoxylin and eosin after fixation and prior to embedding in paraffin wax and sectioning. The choice of fixative is important and best results are obtained using Worcester''s Fluid, a combination of saturated aqueous mercuric chloride, formaldehyde and glacial acetic acid. After fixation, blocks of tissue up to 1.5 cm thick are stained for 7 days in hematoxylin. Excess stain is removed by washing tissues in running water overnight. Tissue blocks are then dehydrated with graded concentrations of ethyl alcohols to 80% and counterstained, with further dehydration, in 0.5% spirit soluble eosin in 90% ethyl alcohol for 5 days. The tissue is subsequently transferred to 90% ethyl alcohol overnight to differentiate eosin staining; dehydration is completed in absolute ethyl alcohol. The blocks are cleared in cedarwood oil and briefly in xylene prior to embedding, sectioning and mounting. Following removal of wax by xylene, coverslips are applied. General morphological and histological features were particularly well differentiated and very selectively and reliably stained by this method. [The tissues used included cat soft palate, monkey pyloric canal, monkey femur, human aorta, and koala bladder.].

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