Dewaxed and hydrated sections were treated with a 0.5% acriflavine hydrochloric acid solution of pH 1.5 for 30 min and then washed with distilled water followed by a rinse in descending concentration of ethanol and again in water. The sections were next treated with a 0.1% alcian blue acetic acid solution of pH 2.5 for 2-20 min, rinsed in water, dehydrated with ethanol and then cleared and mounted. Cartilage matrix appears bluish yellow and lacunar borders and chondrocyte cytoplasm yellowish blue. Mast cell cytoplasmic constituents appear bluish orange in contrast to the pale yellow of the nucleus.
Mitochondrial activity of the indoxyl acetate esterase was proved in absorptive cells of rat jejunum and in parenchymal cells of rat liver by an indoxyl acetate esterase stain. And continuities between the endoplasmic reticulum and the mitochondria were observed by chance in these cells.
Indoxyl acetate esterase was demonstrated in rat liver by an azo-indoxyl method at the fine structural level. The present study on the intracellular localization of esterase activity suggests that the esterase activity in centrifugal fractions might not be the contamination of the microsomal fraction. 10-3 M sodium fluoride completely suppressed the staining of the cytoplasmic membranous structure, but did not suppress the stain of the droplets, exhibiting the so-called peribiliary activity of type A esterase.
In the Prussian Blue method for the histochemical demonstration of iron, some parts of produced pigment are considerably washed away during the dyeing process, because the pigment is water soluble. Consequently decrease and diffusion of the pigment occur and staining of the surrounding tissues take place. The color reaction was carried out in 70% acetone solution in order to decrease the solubility of the pigment. Obtaining the sharp and clear demarcation, this method was applied successfully to the colloidal iron reaction of acid mucopolysaccharides and very strong and clear reaction was obtained.