Henry James Werner

Chromatography and Biological Stains: III. Comparison of the Fat Staining Efficiency of Fractions of Commercial Sudan III Separated by Column Chromatography

Fractions of commercial Sudan III which were separated by column chromatography were compared for fat staining efficiency. Paraffin sections of chromated mouse liver tissue and sections of both fresh and formalin-fixed rat liver tissue or Musca domestica larvae, cut with the freezing microtome, were used. Evidence is presented that a sample of very highly purified Sudan III has no ability to render a fat stain in fresh, formalin-fixed or chromated tissue. However, certain other fractions from the commercial sample, some completely devoid of Sudan III, had good staining characteristics. It is concluded that some substance or substances, other than Sudan III, is responsible for the staining action of the commercial dye.

Morphologic Aspects of the Chest Gland of the Bat, Molossus Ater

The single oval chest gland of the bat, Molossus ater , is situated in the midventral axis at the base of the neck. The gland has a vascular fibrous capsule. Septa extend inward, forming a delicate supporting stroma for the ducts and glandular parenchyma. Two types of glands are present: sebaceous glands which form the main bulk and are distributed throughout the entire structure and sweat glands confined to the innermost portions where they are present in clusters. Ducts of both glands open into a pocketlike integumental fold at the ventral surface.

Morphological Aspects of the Salivary Glands of Nutria

The salivary glands of nutria, Myocaster coypus , consist of the parotid, submaxillary and sublingual major and minor. The first two are pure serous glands. The sublingual major and minor are mixed, having mucous and serous secretory end pieces or mucous alveoli capped by serous demilunes. All the glands have connective tissue capsules consisting mainly of collagenous fibers with a few elastic fibers. Inward projections of the capsule divide the gland into lobes and lobules and surround individual end pieces. The duct systems of all glands are histologically similar.

Chromatography and Biological Stains IV. Preparation of a Suitable Fat Stain from Commercial Sudan III

A variety of modifications of the Bensley and Bensley method for the preparation of a fat-staining solution from commercial Sudan III were examined for dye content by paper chromatography and for fat staining efficiency on frozen sections of Musca domestica larvae. These modifications have led us to believe that the most effective staining agent in commercial Sudan III is a bright orange compound with an Rf value of 0.92 in an isooctane system. This compound could be obtained in good yields by reflux extraction of the commercial dye widi dry petroleum ether (1 g. dye to 100 ml. solvent) for 2 hours, filtering, concentrating to 25 ml., and adding the concentrated solution to a 25 × 200 mm. Celite-silicic acid column. The chromatogram was then developed with petroleum ether for 3 hours, dried by suction and extruded. The orange band was sectioned, air dried and stored. One gram of this dry powder extracted with 10 ml. of 40% alcohol gave an efficient staining solution.