Mallory's Connective Tissue Stain Research Articles

Autographic study of fibrinolytic activity of rat oral epithelium

Fibrinolysis occurred over incompletely keratinized epithelium, as shown by Mallory connective tissue stain; fibrinolysis never occurred over completely keratinized epithelium such as the hard palate and gingiva. Both saline and potassium thiocyanate extracts of incompletely keratinized epithelium showed fibrinolytic activity when examined by the fibrin plate technique; although potassium thiocyanate extracts of completely keratinized hard palate epithelium showed fibrinolytic activity, saline extracts showed none. The results indicate that fibrinolytic autography only demonstrates a saline-extractable activator which is absent or present in relatively small amounts in completely keratinized epithelium.

A histochemical and fine structural study of early extracellular connective tissue in the chick embryo.

AbstractThe histochemistry of developing connective tissues and its relation to early connective tissue fibrils was investigated in the perinotochordal area of the chick embryo. Chick embryos were sacrificed at one, two, three, four, six and ten days of incubation and were prepared for both light and electron microscopy. Five histochemical stains (PAS, alcian blue, Hale colloidal iron, metachromatic toluidine blue, methenamine silver) were used to demonstrate polysaccharides, mucoproteins and mucopolysaccharides. Mature connective tissue elements were demonstrated by Mallory's connective tissue stain and Weigert's resorcin‐fuchsin stain. Routine electron microscopic techniques served to demonstrate extracellular connective tissue fibrils.The first positive response for all histochemical stains occurs on the third day of incubation. Moderate microfibrillar growth precedes this by one day. Light microscopic staining patterns differ from electron microscopic fibrillar arrangements. In the perinotochordal area microfibrils later contribute to the cartilaginous model of the future vertebral body. By the sixth day, dense microfibrillar concentrations appear in the precartilage area where acid mucopolysaccharides are intensely concentrated. Staining for mature connective tissue fibrils does not occur until the tenth day.

THE RELATION BETWEEN THE STAINING PROPERTIES OF THE THYROIDAL COLLOID AND ITS IODINE CONTENT

A new staining technique has been developed for the thyroid gland involving the use of two components of the Mallory connective tissue stain, aniline blue and orange G in reversed proportions.Various indices such as incorporation of radioiodine, epithelial cell height, and number of blue and yellow staining follicles and total number of follicles have been used to test the validity of the color reaction in the colloid. The comparison of these diverse indices strongly suggests that the colloid material which stains with aniline blue corresponds to iodinated thyroglobulin, while the yellow staining material appears to be devoid of biologically active iodinated amino acids.

THE RELATION BETWEEN THE STAINING PROPERTIES OF THE THYROIDAL COLLOID AND ITS IODINE CONTENT

A new staining technique has been developed for the thyroid gland involving the use of two components of the Mallory connective tissue stain, aniline blue and orange G in reversed proportions.Various indices such as incorporation of radioiodine, epithelial cell height, and number of blue and yellow staining follicles and total number of follicles have been used to test the validity of the color reaction in the colloid. The comparison of these diverse indices strongly suggests that the colloid material which stains with aniline blue corresponds to iodinated thyroglobulin, while the yellow staining material appears to be devoid of biologically active iodinated amino acids.

The dermal-epidermal junction; a preliminary study with periodic acid Schiff stain.

The presence or absence of a basement membrane at the epidermal-dermal junction has been, and probably will remain, problematical. That a periodic-acid-Schiff (PAS)-positive zone occurs at the epidermal-dermal junction is undoubted; whether this actually is a basement membrane is questionable. This zone, or membrane, definitely has some characteristics of a basement membrane. The purpose of this project was to study the PAS-stained membrane in a number of skin lesions and was an attempt to determine whether it was truly a basement membrane and even if not, whether its character, its presence, or absence could be of any use in diagnostic histopathology of the skin. Herxheimer, in 1916, using Mallory's connective tissue stain, studied the problem of the epidermal basement membrane and concluded that there was present, immediately below the basal cells of the epidermis, a thin fiber 0.5μ in diameter which was morphologically

A differential staining method for elastic fibers, collagenic fibers, and keratin.

A method allowing for the differential presentation of elastic fibers, other connective tissue fibers, epithelial and other types of cytoplasm, and keratin is described. The procedure is based on the affinity of orcein for elastic fibers, of anilin blue for collagenic material, and of orange G for keratin. Bouin-fixed, tissue-mat embedded sections are stained in Pinkus' acid orcein for 1 1/2 hours and rinsed in distilled water. The sections are differentiated in 50% alcohol containing 1% hydrochloric acid, washed in tap and then in distilled water. The sections are next transferred for I to 2 minutes to the anilin blue, orange G, phosphomolybdic acid combination known as solution No. 2 of Mallory's connective tissue stain, diluted 1:1 with distilled water. They are then rinsed in distilled water, quickly passed into 95% alcohol, and dehydrated in absolute alcohol containing some orange G, after which they are cleared and mounted. Within less than two hours sections may be stained and mounted with the foll...

Histological Aspects of the Glands of the Bat, Tadarida cynocephala (LeConte)

The presence of both glands and glandular tissue in various parts of the bodies of North American bats has been mentioned by several investigators. With the exception of the work of Harrison and Davies (Proc. Zool. Soc. London, 119:331357, 1949), on the facial glands of the European bat, Noctula noctula, studies of a histological nature to determine the type and the probable functions of such tissues seem to be lacking. The work of Harrison and Davies prompted us to begin a study of the glands of some North American bats. The form reported on here, Tadarida cynocephala, is a member of the family Molossidae. A few salient differences have been observed in the glands of the face of this species, as compared with Harrison's and Davies' description of the facial glands of Noctula noctula, which belongs to the family Vespertilionidae. Materials and methods.-Specimens, including both males and females, were collected in Baton Rouge, Louisiana. Tissues were fixed in either Bouin's or Aceto-Zenker's solution depending upon the subsequent stain. All the tissues were dehydrated by means of various concentrations of ethyl alcohol, cleared in cedar oil, and embedded in paraffin. Serial sections were cut at five microns and stained as follows: Harris' haematoxylin with an alcoholic eosin counter stain, Heidenhain's modification of Mallory's connective tissue stain (Azan), Heidenhain's iron haematoxylin, the Feulgen reaction, and Lillie's iron haematoxylin and metachromatic dyes. The first stain is conducive to an over-all examination of the tissues. The azan method was used for identification of connective tissue patterns. The third and the fourth methods were of value because they are, respectively, specific for nuclear patterns and chromatin material. Lillie's metachromatic dye-method was used to detect the presence or absence of mucin within the cells. As a check of the effectiveness of this dye, cartilage was always stained simultaneously, along with the tissue. Discussion.-Two types of facial glands were found, namely, sebaceous and salivary glands. The former are holocrine glands, the secretion of which is derived from the degeneration of the cells of the gland itself, and is eventually expelled onto the skin of the bat. Harrison and Davies (loc. cit.) separated these glands in Noctula noctula on the basis of their location into pararhinal glands (on the muzzle) and submental glands (beneath the lower jaw). Glands similar to these were found in the corresponding body regions of Tadarida cynocephala, but they were also found elsewhere on the body of the bats. These glands are the etiology of the strong, unpleasant odor and the oily pelage of Tadarida. The salivary glands consist of three distinct pairs of glandular masses. One pair is located in the muzzle, above the upper jaw, and empties by means of a large duct into the inner side of the upper jaw, distal to the mandible. The second pair of glandular masses, located in the lower jaw, empties onto the mandibu395

An Easily Controlled Regressive Trichromic Staining Method

Three modifications of Mallory's connective tissue stain are described and some features of the action of picric acid are discussed.In the first and most critical method the nuclei are stained in an iron hematoxylin and then differentiated in a picric acid solution containing orange G. This not only differentiates the nuclei, but stains all other elements yellow. The section is then washed in running water to remove the yellow color from all tissues except those which are to remain yellow in the final preparation (usually the erythrocytes). The section is next stained in an acid fuchsin mixture and then differentiated until the desired depth and contrast is obtained. Staining in anilin blue follows and this in turn is differentiated to suit. The section is then dehydrated and mounted.In the second method the nuclei are stained in hemalum (e.g. Harris's) for a short time; the section is then rinsed and immersed in a mixture of picric acid and acid fuchsin and thereafter is differentiated; it is next passed...

PHYSIOLOGICAL SIGNIFICANCE AND MORPHOLOGY OF THE CARMINE CELL IN THE CAT'S ANTERIOR PITUITARY1

IN A STUDY of the anterior pituitary of the female rabbit (1, 2) it was found that striking changes occurred in the composition of the acidophil cells after mating. This reaction was demonstrated by means of a method which separated the acidophils into two distinct tinctorial types (3). The glands were fixed in sublimateformalin followed by an adaptation of Heidenhain's azan modification of Mallory's connective tissue stain. The ordinary or standard acidophil was colored yellow with orange G. Other acidophil cells which become conspicuous during sexual activity were characterized by the coarseness and refractivity of their granules, and took a brilliant red stain due to their special affinity for azocarmine. The chromophobes were not colored or appeared a light pink, and the basophils stained blue with analinblue. In estrous rabbits the modified acidophils (carmine cells) are usually small and inconspicuous, but after mating they increase in size and number and become

Reversibility of Hyalinization in the Mouse Uterus Produced by Injections of Estrogen, and the Changes in the Mammary Gland and Ovaries after Cessation of Injections

In former investigations (1) we observed the deposition of a hyaline substance in the connective tissue of the uterine mucosa and muscularis of mice injected with large doses of estrogen for a period of two months or longer. This hyaline material acted in certain respects like a foreign body, in some instances causing the formation of epithelioid and small giant cells which helped in its absorption as well as in its invasion and replacement by connective tissue. Absorption was accompanied or followed, however, by the deposition of new hyaline, and in this way the abnormal condition of the uterine connective-tissue stroma and muscularis was perpetuated. There was evident a tendency of the hyaline to persist, especially at the borderlines separating different structures, such as blood vessels and glands, from the surrounding stroma. In the uterine cervix conditions were similar to those observed in the uterus, but in certain parts of the vagina, especially around the vaginal folds and between folds and cervix, a rarefaction of the tissue took place, apparently due to the occurrence of liquefaction processes in the hyaline material or to its deposition in a diluted form. As to the chemical nature of the substance deposited in the uterus, cervix, and vagina, the reactions for amyloid were negative; on the other hand, it stained strongly with eosin. In further tests it was found to stain red with Van Gieson stain and strongly blue with Mallory's connective-tissue stain. It did not take Weigert's stain for fibrin. In these reactions the behavior of the hyaline substances deposited in the uterus and in certain parts of the cervix and vagina of the mouse following injections of large doses of estrogen is identical with that of collagen.

Further Experiments with the Masson Trichrome Modification of Mallory's Connective Tissue Stain

Several dyes, notably ponceau 2R, azofuchsin 3B, nitrazine yellow, and Biebrich scarlet may replace imported “ponceau de xylidin” in the Masson ponceau acid fuchsin mixture. Of these Biebrich scarlet appears to be the best and may be used without acid fuchsin.A mixture of equal parts of 5% solutions of phosphomolybdic and phosphotungstic acids is much superior to either acid alone and gives adequate mordanting in 1 minute at 22°C.With the fast green modification, times in plasma and fiber stains can be reduced to 2 minutes each. With anilin blue a 4-minute plasma stain is required. One-minute final differentiation in 1% acetic acid is adequate.Primary mordanting of formalin material may be accomplished by 5 minutes in saturated aqueous mercuric chloride or 2 minutes in saturated alcoholic picric acid. Three minutes washing in running water is required after these mordants.

Differentiation of the Secretory Cells of the Pars Nervosa of the Hypophysis Cerebri

A method was found by means of which two types of granular cells in the pars nervosa of the hypophysis cerebri could be preserved in permanent preparations so as to retain the appearance these cells presented in fresh tissue and in tissue cultures. The essential points of the technic were the fixation for 24 hours of the hypophysis in a solution composed of 2 parts of 3% potassium bichromate plus one part of a one-half saturated solution of bichloride of mercury in 95% alcohol. Sections of this material were prepared using dioxan in place of the higher alcohols and xylene. The sections were stained by means of Mallory's connective tissue stain leaving the sections in the fuchsin solution for 30 minutes and in the mixture of aniline blue, orange G and phosphotungstic acid for 1 to 24 hours.

Differentiation of Two Classes of Acidophiles in the Anterior Pituitary of the Female Rabbit and Cat

A differential stain for the anterior pituitary of mammals, based directly on Heidenhain's ‘azan’ modification of Mallory's connective tissue stain has been devised. Tissue is fixed for 24 hours in a saturated solution of corrosive sublimate in physiological saline (90 parts) and formalin (10 parts) and washed directly in 70% alcohol for 48 hours. Sections are treated on the slide with a 3% solution of potassium bichromate for 12 hours. Two classes of acidophiles are demonstrated: one which stains selectively with azocarmine; and the ordinary acidophile which stains with orange G. The special acidophile has been demonstrated in the female rabbit and cat but has not been found in the mouse or rat.

A modification of Mallory's connective tissue stain with a discussion of the principles involved

A modification of Mallory's connective tissue stain with a discussion of the principles involved

The histology of the paraphysis of Amblystoma

AbstractThe paraphysis of adult Amblystoma is made up of low columnar ependymal epithelit m which forms the paraphyseal tubules which end blindly and which communicate with one another by a common mouth with the third ventricle. Between the paraphyseal tubules venous sinusoids anastomose freely with one another forming a complicated rete. The sinusoids are made up entirely of endothelium. The blood supply to the paraphysis is entirely venous.Mitochondria were found in great abundance in the paraphysis of one female just previous to laying. Other specimens showed very few present. No conclusions can be drawn from these few observations as to the relationship between physiological activity and cellular structures.The Golgi apparatus was observed definitely localized between the nucleus and the ventricular end of the cell.Many large crystalloids were also observed to be localized between the nucleus and the ventricular end of the cell.Intercellular spaces are readily observed in sections stained with Mallory's connective tissue stain. Nassonow's osmic acid technique for the Golgi apparatus and Benda's crystal violet and alizarin stain clearly bring out the intercellular canals. Acid fuchsin stained particles within the intercellular spaces are more abundant toward the sinusoids than the cavities of the paraphyseal tubules. The intercellular canals have not been seen to communicate with either the sinusoids or the cavities of the paraphyseal tubules in any of the preparations observed.

A study of the collagen of the placenta

A study of placentas of varying ages has been made. The Lee-Brown modification of Mallory's connective tissue stain has been used. The purpose has been to trace the development of collagen. The formation of collagen has been shown to take place at the junction of the fetal and maternal cells. It is not formed in decidua where there are no fetal cells. There is a marked absence of collagen formation in placenta accreta, in tubal pregnancy, and in chorionepithelioma. A definite basement membrane of the villous epithelium has been demonstrated in the latter half of pregnancy. This has a definite width and thickness according to the age of the placenta. In cases of eclampsia and severe preeclamptic toxemia, a definite thickening of the basement membrane has been demonstrated which is not found in the normal ripe placenta, in nephritis, or in mild toxemias. A study of placentas of varying ages has been made. The Lee-Brown modification of Mallory's connective tissue stain has been used. The purpose has been to trace the development of collagen. The formation of collagen has been shown to take place at the junction of the fetal and maternal cells. It is not formed in decidua where there are no fetal cells. There is a marked absence of collagen formation in placenta accreta, in tubal pregnancy, and in chorionepithelioma. A definite basement membrane of the villous epithelium has been demonstrated in the latter half of pregnancy. This has a definite width and thickness according to the age of the placenta. In cases of eclampsia and severe preeclamptic toxemia, a definite thickening of the basement membrane has been demonstrated which is not found in the normal ripe placenta, in nephritis, or in mild toxemias.

The Membrana Granulosa of the Mouse

THE accompanying figure (Fig. 1) illustrates the membrana granulosa of the developing follicle of the mouse, from a preparation fixed with chromic acid and osmium tetroxide and stained with Mallory's connective tissue stain. As the darkly staining cells shown in particular at S.C. are not commonly stressed in descriptions of this tissue1,2, apparently because their presence is regarded as symptomatic of degeneration, it is desired to direct attention to some of their peculiarities.

Mallory's Connective Tissue Stain Following Hematoxylin

The following combination of hematoxylin with Mallory's connective tissue stain is useful in bringing out nuclei as well as in differentiating tissue:Slightly overstain in Mayer's hematoxylin (50 g. potassium alum and 0.2 g. sodium iodate added to 1 liter 0.1% aqueous hematoxylin). Wash; and stain 30 seconds to 1 minute in 0.04% aqueous acid fuchsin-Stain 4 minutes in: 0.5 g. anilin blue and 2 g. orange G dissolved hi 100 cc. of 1% aqueous phosphomolybdic acid. Pass thru 95% alcohol to absolute; clear in xylol and mount in balsam.

Cell Types and Their Proportion in Pars Anterior of Adult Male Human Hypophysis.

There is considerable disagreement on the number of distinct cell types in the anterior lobe of the hypophysis, as well as on the question of the relative number of each type under normal conditions. As many as 7 different kinds of epithelial cells have been described. How many of these are merely different functional stages of the same cell is not settled; but for practical purposes, at least 3 kinds can be regularly recognized by relatively simple technique. They are: chromophobe (chief, reserve, clear, undifferentiated, neutrophil), acidophil (eosinophil, oxyphil, alpha) and basophil (cyanophil, beta). The tendency seems to be to regard the acidophils as predominating and either the chromophobes or basophils as the fewest in number, although a few authors estimate that the chromophobes are the most numerous. An increase in the relative number of basophils in old age is also a widespread notion. It is obviously of prime importance to settle these questions in order to properly interpret abnormalities in the hypophysis, such as are now being reported frequently in the pathological literature. This is a preliminary report on a differential count of the cells in the anterior lobe of 100 human hypophyses from normal male adults, according to the method outlined in a previous communication. The only change introduced since then is to put the section first for about one-half minute in ordinary hematoxylin and washing well with distilled water before staining with Mallory's connective-tissue stain. Only cases of sudden and usually accidental death, in which the hypophysis was fixed within 12 hours after death, were used. They ranged in age from 18 to 78 years. The chromophobes represent on an average 52% of the total cells (from 34% to 66%). The scarcity of chromophobes, so often mentioned, is not substantiated, since they are normally never below one-third of all the cells. They are also the least variable, the coefficient of variation being 15. Many of these cells contain very little cytoplasm so that they do not appear as numerous as an actual count indicates.

THE VALUE OF MALLORY'S CONNECTIVE TISSUE STAIN FOR THE DEMONSTRATION OF VARIATION IN THYROID COLLOID

1. Mallory's connective tissue stain used in connection with fixation of tissue by Zenker's fluid or bichloride of mercury demonstrates the presence or absence of iodin in the thyroid colloid, as is evidenced by parallel chemical determinations, iodin feeding, and test-tube experiments. 2. Greatly increased activity of the heart is accompanied by a loss of thyro-iodin from the colloid of the thyroid. 3. This method, while apparently not adaptable to clinical use, may prove of value in further experimental work on the thyroid.