A SIMPLE METHOD FOR THE STAINING OF VIRAL INCLUSION BODIES WITH FUCHSIN S AND HEMATOXYLIN

Abstract

A simple method for the staining of viral inclusion bodies was described.
Procedure:
(1) Fix in CARNOY's fluid, 10% formalin or absolute alcohol.
(2) Prepare paraffin sections in routine.
(3) Stain sections in 0.5% fuchsin S for 5 minutes.
(4) Wash off exess reagant in water for a moment.
(5) Treat with 1% phosphowolframic acid in destilled water for 20 minutes.
(6) Wash in water for a moment.
(7) Place in MAYER's hemalaum (hematoxylin) for 5 minutes.
(8) Wash in running water for 10 minutes.
(9) Dehydrate, clear and mount as usual.
Smear preparations can also be stained, but mount in Canada balsam or examine under microscope with oil immersion lens.
The ground substance of type A intracytoplasmic inclusion body (MARCHAL body) appeared deep scarlet in EHRLICH, YOSHIDA and sarcoma 180 ascites tumor cells, the chorioallantoic membrane of chick embryo, EHRLICH ascites tumor cells transplanted onto C. A. M., and the liver, other organs and tissues of mouse infected with ectromelia virus. The elementary bodies and the outline of type A intracytoplasmic inclusion body (BOLLINGER body) appeared deep scarlet in the C. A. M. of chick embryo and the skin of chick infected with fowlpox virus. The ground substance of this inclusion body was stained with neither fuchsin S nor hematoxylin. The type B intracytoplasmic inclusion bodies of these two kinds of viruses were not distinguished from the cytoplasm with this staining method. The classical type A intranuclear inclusion bodies appeared scarlet in the bronchial epithelium of monkey infected with measles virus and in the C. A. M. and the liver of chick embryo infected with herpes simplex virus. The inhalt of swollen nucleus infected with herpes simplex virus was stained slightly purple. Red blood cells and granules of eosinophilic leucocytes were stained deep scarlet. Nucleoli, some parts of degenerated cytoplasm and some of degenerated nuclei were also stained scarlet. Nuclei were stained deep blue. All others appeared faint purple.
By means of this staining the acidophilic inclusion bodies could be easily perceived.