A rapid technique for alkaline phosphatase enzyme activity in tissues.

A rapid method is described for serial determination of alkaline phosphatase enzyme activity in the rat kidney and the developing avian vertebra by using an improved paraffin embedding process and a slide staining rack. Tissue samples fixed 12 to 24 h in 80% ethanol at 0 to 4 degrees C are dehydrated in absolute acetone at 0 to 4 degrees C, and then embedded in paraffin of 50 degrees C via benzene at room temperature. The alkaline phosphatase enzyme activity is essentially demonstrated with the method of GOMORI and TAKAMATSU but modified for the use of the slide staining rack. With the new technique, the enzyme activity is completely preserved and reproducible for years in the paraffin embedded tissues. The good quality of the sections enables quantitative evaluations by video scanning.

An inexpensive, easily constructed slide staining rack.

An inexpensive, easily constructed slide staining rack is described that has been successfully used for staining histological sections and various smears. The major advantages offered by the rack are: rapidity, good quality, and decreased exposure of the technicians to solvents. To improve staining capacity and quality of slides, a simple mechanical slide staining rack has been developed in our laboratory. Here, construction, operation and evaluation of this device are described.

Pathological features of 3-chloro-4-methyl benzamine HCl toxicity in rooks (Corvus frugilegus L.) and pheasants (Phasianus colchicus L.).

Gross and microscopic changes in response to single po. doses of avicide 3-chloro-4-methyl benzamine HCl were studied in rooks (Corvus frugilegus L.) and pheasants (Phasianus colchicus L.). The observed changes followed a dose- and survival time related pattern and rooks were considerable more susceptible than pheasants to the toxicity of the compound. No effect was found in rooks and pheasants which survived safely 0.7 to 1.0 and 7.0 to 10.0 mg/kg doses, respectively. Early acute effect, in rooks died from 7.0 to 16.0 mg/kg within 16 to 18 h, included: congestion of the major organs, fatty degeneration of the liver, and extensive parenchymal degeneration of the kidney, particularly the proximal tubular epithelial cells. Progressive late effect developed in rooks and pheasants at 2.2 to 5.0 and 16.0 to 120.0 mg/kg doses, respectively, which died in coma between 32 to 80 h. It was characterized by a grayish white, frost-like material of uric acid overlaying the serosal surfaces of the various organs, accompanied by sterile inflammation and necrosis in the affected and adjacent tissues. Few urate deposition was also seen in some of the kidneys. Infrequently, interstitial pneumonia did also occur. The lesions produced by the toxic effect of the test compound seemed to be analogous to that of avian visceral gout.

Video scanning for the quantitative determination of alkaline phosphatase activity in the rat kidney.

The use of a video scanning equipment (Opton Micro-Videomat) for the quantitative determination of alkaline phosphatase activity in histological sections of the rat kidneys is presented. A photomicroscope with a telecamera, a video monitor, a computer and a recorder are incorporated into the system. Automatic measurement, accelerated detection, accuracy, sensitivity and reproducibility, as well as elimination of time-consuming work involved in other available methods are found to be the main advantage of the method.