scholarly journals ALDEHYDE-FUCHSIN FOR PITUITARY CYTOCHEMISTRY

1959 ◽  
Vol 7 (2) ◽  
pp. 98-100 ◽  
Author(s):  
HERBERT ELFTMAN

The component of the aldehyde-fuchsin stain which is useful for pituitary staining requires more delicate manipulation than the elastic tissue component. Manufacture of the stain for pituitary purposes may he shortened to 26 hours by subjecting a 0.5% basic fuchsin solution in 70% ethanol, 0.75% paraldehyde and 1.25% HCl to incubation at 37°C. The pH of this solution precludes background staining, allowing aldehyde-fuchsin to be followed by the periodic acid-Schiff routine. Aldehyde fuchsin may be rejuvenated by adding basic fuchsin in alcoholic solution. Oxidation of pituitary tissue allows aldehyde-fuchsin to stain not only thyrotrophs but gonadotrophs as well and accentuates the staining of the hypothalamic hormones and the Golgi bodies. Since some fixatives, such as dichromates, oxidize tissue, assay of the significance of aldehyde-fuchsin staining involves awareness of the influence of the particular fixative employed. Spectrophotometric analysis of young and old aldehyde-fuchsin stains supports the assumption that staining of the pituitary thyrotrophs may involve other constituents of the stain complex than those relied on for elastic tissue visualization.

1972 ◽  
Vol 20 (11) ◽  
pp. 896-907 ◽  
Author(s):  
ROGER R. MARKWALD ◽  
WILLIAM N. ADAMS SMITH

Mucosubstances (MS) were examined in 10½-14½-day embryonic rat hearts utilizing nonaqueous fixatives or formaldehyde vapor-fixed frozen sections hydrated in concentrated solutions of cetylpyridinium chloride. Ribonuclease-resistant, polyanionic sites were limited to the extracellular cardiac jelly, endocardium and fibroblastic cells (cushion tissue) associated with the endocardium. The cardiac jelly and endocardium of day 10½ embryos principally contained a hyaluronic acid-like carboxylated mucosubstance whose alcianophilia at pH 2.5 was abolished by hyaluronidase but was resistant to NaOH extraction and neuraminidase and trypsin digestion. A critical electrolyte concentration of 0.2 M MgCl2 abolished alcianophilia. On days 13½-14½ carboxylated MS were restricted to cushion tissue and partially resisted mild methylation. Sulfated MS were limited to primitive endocardial cells which gave origin to cushion tissue. Dye deposits of aldehyde fuchsin, high iron diamine or Alcian Blue (pH 1.0) were localized on cell surfaces and such staining was prevented by strong (60°C) methylation. Hyaluronidase sensitivity of sulfated MS decreased with gestation. The critical electrolyte concentration varied from 0.5-0.7 M MgCl2 on days 11½-12½ to 0.8-0.9 M MgCl2 after day 12½. The sulfated MS of endocardial cells were preceded by a transitory accumulation of diastase-resistant, periodic acid-Schiff-positive material. Possible roles of MS in normal and abnormal cardiac septation processes are discussed.


1956 ◽  
Vol s3-97 (39) ◽  
pp. 379-391
Author(s):  
C. L. FOSTER

Human anterior pituitary tissue that had been removed at operation and immediately fixed was examined by a number of cytological and histochemical methods and by phase contrast and electron microscopy, and compared with similar material obtained post mortem. The general histological picture of good post-mortem material (not more than 4 hours post mortem) compared well with the surgically-removed tissue. For the study of silver impregnations of the Golgi substance, however, material removed at operation was found to be greatly superior. Evidence was obtained showing that the intracellular lipid inclusions seen post mortem were not artifacts resulting from cytolytic changes. There appeared to be no relationship between these lipid bodies and the Golgi material as revealed by the Aoyama method. No unequivocal dimorphism of the Golgi bodies, correlated with α- and β-cells, such as has been reported to occur in certain other mammals, was observed. Phospholipid was present in the granules of a substantial proportion of the α-cells. It was found that most of the cells which had been designated as β-cells after the application of certain routine staining methods, and most of the Gram-positive cells, reacted positively to the Periodic acid Schiff test: these cells could therefore be regarded as true β- or mucoid cells. A method for the demonstration in frozen sections of the cell-types, together with the lipid inclusions, is described.


1973 ◽  
Vol 21 (9) ◽  
pp. 794-803 ◽  
Author(s):  
KAZUYORI YAMADA

The effect of digestion with Streptomyces hyaluronidase upon certain histochemical reactions of hyaluronic acid-containing tissues has been studied in a series of human, mammalian and avian specimens. These histochemical reactions are those for the demonstration of neutral and sulfated and nonsulfated acid mucosaccharides such as periodic acid-Schiff, Alcian Blue (pH 1.0), azure A (pH 1.5), high iron diamine, aldehyde fuchsin, Alcian Blue (pH 2.5), Alcian Blue (pH 2.5)-periodic acid Schiff, azure A (pH 4.5), low iron diamine and colloidal iron. In addition, the effect of digestion with testicular hyaluronidase upon the same reactions of the same tissues was observed for comparison. Digestion with Streptomyces hyaluronidase diminishes the staining reactions due to hyaluronic acid but fails to affect those for neutral and sulfated acid mucosaccharides. These results indicate that digestion with Streptomyces hyaluronidase is a method of choice for the identification of hyaluronic acid in mucosaccharide histochemistry.


Author(s):  
Joanna Klećkowska-Nawrot ◽  
Karolina Goździewska-Harłajczuk ◽  
Renata Nowaczyk ◽  
Krzysztof Krasucki

The aim of the present study was morphological and histochemical analysis of the lacrimalgland (LG) in African black ostrich Struthio camelus domesticus in the embryonic and postnatalperiod. Studies were conducted on 50 ostriches aged between the 28th day of incubation until7 months old. Tissue sections were stained with haematoxylin and eosin, Azan trichrome,periodic acid-Schiff, Alcian blue pH 2.5, aldehyde fuchsin and Hale’s dialysed iron. The LGin ostrich was classified as a tubulo-acinar type. The primordia of the lobes were determinedin the LG structure on the 28th day of incubation, whilst the weakly visible lobes with aciniand tubules were observed on the 40th day of incubation. Morphometric studies of the LGshowed steady growth, characterised by an increase in both length and width. Histometricmeasurements of lobe size showed little difference between the first, second and third agegroups, whilst in the fourth age group a marked increase in size of lobes was observed.The study showed that, apart from morphological changes, during the growth of the LGthe character of acid mucopolysaccharides changed. Sulphated acid mucopolysaccharideswere indicated, particularly with aldehyde fuchsin (AF) staining in the fourth age group.The Hale’s dialysed iron (HDI) staining showed a low concentration of carboxylated acidmucopolysaccharides in the first and second age groups and a higher concentration in thethird and fourth age groups. Periodic acid-Schiff staining (PAS)-positive cells were observedin each age group, but only a small number of cells with a weakly PAS-positive reaction weredemonstrated in the first age group.


1983 ◽  
Vol 29 (5) ◽  
pp. 836-839 ◽  
Author(s):  
S M Trivedi ◽  
C G Frondoza ◽  
R L Humphrey

Abstract We have modified the periodic acid-Schiff staining technique for glycoproteins for use with thin agarose-film electrophoresis membranes. With this procedure, carbohydrate-rich proteins can be detected with negligible background staining and insignificant staining of nonglycoproteins such as albumin and nonglycosylated Bence Jones proteins (free light chains). On the other hand, carbohydrate-rich M components of immunoglobulins M and A in serum and in cerebrospinal fluid from patients with plasma cell dyscrasia are readily detected. The technique is two- to threefold more sensitive than Ponceau S. Glycoproteins in serum and body fluids can be determined as a routine analytical procedure.


1955 ◽  
Vol s3-96 (34) ◽  
pp. 193-201
Author(s):  
E. J. W. BARRINGTON ◽  
A. J. MATTY

Evidence is given for the existence of two main types of cyanophil cell in the median zone of the glandular lobe (adenohypophysis) of the minnow, distinguishable by their distribution and by their cytological characteristics. Both types are positive to the periodic acid Schiff (PAS) technique, but one (type 2 of this account) also gives a positive response to the aldehyde-fuchsin (AF) technique of Gomori, as used by Halmi and by Purves and Griesbach in studies of the mammalian pituitary. In fish which have been immersed in thiouracil solution the type 2 cells show degranulation and vacuolation, and their characteristic positive AF response is very greatly weakened or lost. For these reasons the type 2 cells are believed to be responsible for the secretion of thyrotrophin, and appear to be very closely comparable with the thyrotroph cells of the pituitary of the rat.


1966 ◽  
Vol 14 (2) ◽  
pp. 159-166 ◽  
Author(s):  
A. REES MIDGLEY

Luteinizing hormone (LH) was localized immunohistochemically to cells in the pars anterior and pars tuberalis of the human pituitary gland. Upon restaining with a variety of histochemical procedures, LH-containing cells were found to be aldehyde fuchsin and periodic acid-Schiff (PAS) positive; however, the intensity of histochemical staining failed to correlate with that of fluorescence. Only a few of the PAS- or aldehyde fuchsin-positive cells contained LH. In contrast to the above, with the performic acid-alcian blue-PAS-organge G sequence of Adams, all cells containing LH were turquoise colored, alcian blue-positive (S1 mucoid cells) and vice versa. This study was performed with antiserum specific for human chorionic gonadotropin which was shown to cross react in immunodiffusion analysis with purified human pituitary luteinizing hormone and to be capable of neutralizing its biological activity.


1985 ◽  
Vol 33 (4) ◽  
pp. 287-294 ◽  
Author(s):  
J C Fanning ◽  
E G Cleary

The microfibrils associated with elastic tissue have been shown to be predominantly proteinaceous. On the basis of their affinity for cationic stains, including ruthenium red, they have been assumed to be glycoprotein, but more evidence to support this claim has not been adduced. Despite repeated investigation of glycoprotein materials obtained by extraction of elastic tissues with reagents that appear to remove microfibrils, the chemical composition of elastin-associated microfibrils remains obscure. An electron microscopic study of the microfibrils in two elastin-rich tissues (bovine nuchal ligament and aorta) during their development was pursued using more specific histochemical methods. The periodic acid-alkaline bismuth stain (analogous to the periodic acid-Schiff stain for glycoproteins in light microscopy) has been adapted for this study. Specific aldehyde groups (confirmed by blocking with m-aminophenol or sodium borohydride) were identified after periodate oxidation as fine granules of bismuth stain. These were shown to localize specifically along the elastin-associated microfibrils in a finely punctate form. Staining of the amorphous elastic component did not occur except for a fine rim adjacent to the microfibrils. Lectin binding with concanavalin A (with ferritin markers) confirmed that there are glucose- or mannose-containing proteins associated with the microfibrillar component of elastic tissue. This was true of these microfibrils in all layers of the aortic wall and throughout the ligament. It was also true of mature adult tissues in which there was a lesser proportion of microfibrils. It is concluded that elastin-associated microfibrils really are associated with glycoprotein(s).


Author(s):  
E. Akat ◽  
H. Arıkan ◽  
B. Göçmen

This study was carried out to assess the localization of hyaluronic acid (HA) and the distribution of glycoproteins in the gastrointestinal system of adult Hyla orientalis. Histochemical analysis of the gastrointestinal system in H. orientalis showed that mucous content included glycogene and/or oxidable dioles [periodic acid/Schiff (PAS)+], neutral or acid-rich (PAS/AB pH 2.5+), sialic acid residues (KOH/PAS+) and acid sulphate [Aldehyde fuchsin (AF)+] glycoproteins. However the mucus content was not the same in stomach, small and large intestine. The mucus content of stomach included only glycogene and/or oxidable dioles and sialic acid residues. Besides these histochemical methods, the localization of HA was detected using biotinylated hyaluronic acid binding protein labeled with streptavidin-fluorescein isothiocyanate (FITC). In the extracellular matrix of the submucosa, the reaction for HA was evident. Since HA was located in submucosa beneath the epithelial layer of gastrointestinal system, it has a significant role in hydric balance, and essential to provide the gastrointestinal system integrity and functionality. According to biometric results, there were statistical differences between small and large intestine in terms of the amount of material stained positive with PAS/AB, PAS, KOH/PAS and AF/AB. Additionally, number of goblet cells in the small and large intestine was significantly different.


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