Periodic acid-thiocarbohydrazide-silver methenamine(PATS) reaction: An improved silver methodology for light and electron microscopy

1984 ◽  
Vol 42 ◽  
pp. 264-265 ◽  
Author(s):  
B. Giammara ◽  
T. Romaine ◽  
W. Ambrose ◽  
J. Hanker
Keyword(s):  
Electron Microscopy ◽  
Osmium Tetroxide ◽  
Periodic Acid ◽  
Patent Application ◽  
Light And Electron Microscopy ◽  
Basement Membranes ◽  
Full Description ◽  
Periodic Acid Schiff ◽  
Reticular Fibers ◽  
Pas Reaction

Many variations of the periodic acid-Schiff(PAS) reaction have been utilized for electron microscopy based on the Gomori periodic acid-silver methenamine reaction (1) or the periodic acid-thiocarbohydrazide-osmium tetroxide(PATCO) reaction (2,3). These reactions are widely employed and have been very useful for the demonstration of one or more biomacromolecules or structures such as glycogen, basement membranes, reticular fibers or lipopolysaccharide. However, these reactions have various drawbacks such as complexity of methodology, ability to stain only a limited number of these components, or lack of adaptability for both light and electron microscopy. Our newly devised PATS reaction is relatively easy to perform. A full description of the details must await the outcome of a pending patent application. It consists essentially of a stepwise treatment of the sample with periodic acid, thiocarbohydrazide(TCH) and silver methenamine.

scholarly journals A MORPHOLOGIC AND CYTOCHEMICAL STUDY ON THE GREAT ALVEOLAR CELL

1966 ◽  
Vol 14 (12) ◽  
pp. 884-897 ◽  
Author(s):  
SERGEI P. SOROKIN
Keyword(s):  
Electron Microscopy ◽  
Periodic Acid ◽  
Light And Electron Microscopy ◽  
Multivesicular Bodies ◽  
Alveolar Cells ◽  
Periodic Acid Schiff ◽  
Alveolar Cell ◽  
Cytochemical Study ◽  
Lysosomal System ◽  
Chloroform Methanol

Lungs from marsupials, bats and rodents were studied by light and electron microscopy. In all three groups, the great alveolar cells exhibit similar morphologic and cytochemical characteristics. Cytoplasmic vacuoles seen in these cells by light microscopy correspond to cytosomes that are demonstrable in them by electron microscopy. Such cytosomes are osmiophilic, periodic acid-Schiff-positive and stainable with Sudan black after acetone extraction. After fixation in a mixture of aldehydes, followed by extraction in chloroform-methanol and postfixation in osmium tetroxide, cytosomes lose their osmiophilia. The cytoplasm of the great alveolar cell is notable for a loosely ordered granular endoplasmic reticulum, an extensive Golgi apparatus and numerous multivesicular bodies. Many forms transitional in appearance between multivesicular bodies and cytosomes are present. In these, osmiophilic matter occupies the intervesicular space. It is proposed that these bodies are the precursors of cytosomes. The cytosomes are interpreted to be products of the "lysosomal" system in this cell. Ultimately they are secreted onto the alveolar surface.

Effects of O3 on submucosal gland of bonnet monkey trachea

1983 ◽  
Vol 41 ◽  
pp. 678-679
Author(s):  
S. Yamashiro ◽  
D. Wilson ◽  
J. St. George ◽  
D. Hyde ◽  
C. Plopper ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Periodic Acid ◽  
Light And Electron Microscopy ◽  
Lung Parenchyma ◽  
Uranyl Acetate ◽  
Upper Airways ◽  
Periodic Acid Schiff ◽  
Transmission Electron ◽  
Bonnet Monkey ◽  
Submucosal Glands

In the past, ozone inhalation studies have focused on the lower airways and lung parenchyma. The purpose of this study was to evaluate the effects of ozone on submucosal glands of upper airways. Six adult male bonnet monkeys were exposed to 0.64 ppm ozone continuously for 7 days, and three were exposed to chamber conditions without ozone. The animals were exsanguinated under barbiturate anesthesia. The trachea and lung were fixed by airway infusion of Karnovsky's fixative, which was adjusted to pH 7.4 and 440 milliOsmols. Sagittal sections of ventral trachea were embedded in glycol methacrylate and Araldite 502 for light and electron microscopy. One micrometer methacrylate sections were stained with Alcian blue-periodic acid Schiff (AB/PAS). Selected areas of Araldite-embedded tissue were sectioned for transmission electron microscopy, stained with uranyl acetate and lead citrate and examined with a Zeiss EM 10. Volume percentages of the lumen, granular and nongranular regions of fhe gland and the duct wall, respectively, were estimated by stereologic methods on AB/PAS stained sections.

Positive stain for light and electron microscopic demonstration of spirochetes in subgingival plaque and crevicular fluid samples from periodontal diseased sites

1989 ◽  
Vol 47 ◽  
pp. 1082-1083 ◽  
Author(s):  
B. Giammara ◽  
E.J. Burkes ◽  
R. Scruggs ◽  
G. Greco ◽  
P. Yates ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Periodic Acid ◽  
Light And Electron Microscopy ◽  
Subgingival Plaque ◽  
Microbial Count ◽  
Electron Microscopic ◽  
Microbial Composition ◽  
Periodic Acid Schiff ◽  
Clear Cut ◽  
Crevicular Fluid

In a recent study of 400 subgingival plaque samples from over 110 adult periodontitis patients, spirochetes were the overwhelming microbial type, averaging about 45% of the microbial count. This finding supports earlier arguments that spirochetes are pathognomonic in periodontal disease. Other studies had shown clear-cut differences in the microbial composition of healthy and diseased subgingival sites — the proportion of spirochetes being significantly higher in the latter. Another study indicated that periodontal deterioration at these sites could be predicted better by increased proportions of motile rods and spirochetes than by clinical measurements. However, spirochetes of all sizes and species do not show the same degree of association with periodontal breakdown. Moreover, spirochetes are usually difficult to culture and stain; they are generally monitored by darkfield or phase contrast microscopy.The PATS reaction, a modified periodic acid-Schiff(PAS) reaction which deposits silver for light and electron microscopy appears to stain Gram(-) bacteria positively as well as neutrophils and activated macrophages. When studying the stained Gram(-) bacteria on coverslip smears of subgingival plaque or crevicular fluid samples of patients by light microscopy, varying numbers of intensely stained spirochetes of different sizes were observed (Figs. 1,2). More spirochetes were usually seen in samples from diseased sites. After drying replicate PATS-stained coverslips with hexamethyldisilazane they were sputter coated with gold, and. then examined by the SEI and BEI modes of scanning electron microscopy (Figs. 3-6). A permanent record of the proportions of large, medium and small spirochetes at each site could thus be obtained. Generally, greater numbers of gram negative bacteria including some spirochetes were stained in samples from diseased sites. At some sites, however, spirochetes were the predominant microbes in both crevicular fluid and subgingival plaque (Fig. 1).

Banded Structure in a Hepatocellular Carcinoma

1977 ◽  
Vol 35 ◽  
pp. 510-511
Author(s):  
E. C. Chew ◽  
W. C. Chan
Keyword(s):  
Electron Microscopy ◽  
Hepatocellular Carcinoma ◽  
Osmium Tetroxide ◽  
Periodic Acid ◽  
Uranyl Acetate ◽  
Lead Citrate ◽  
Periodic Acid Schiff ◽  
Cacodylate Buffer ◽  
Tissue Area ◽  
Neural Tumor

Extracellular banded structures were first reported by Luse (1) in a neural tumor and subsequently by others in many types of tissues. This subject was summerized in detail by Sun and White in 1975 (2). This communication reports observations of banded structures discovered by electron microscopy in the study of a human hepatocellular carcinoma. The tissue was fixed in 3% glutaraldehyde in 0.1 M cacodylate buffer and post-fixed in buffered osmium tetroxide. Sections were stained with uranyl acetate and lead citrate. Thick sections, about 1 - 2 μ, were stained with periodic acid-Schiff's reagent involving heating of the slides.Banded structures are observed in the connective tissue area intermingled with collagen fibrils and are usually fusiform in shape (Fig. 1). The fusiform bodies average 0.5 μ in diameter and are outlined with periodicity of 800 to 1000 A°. Each period consists of a light and a dark band. Fine filaments of about 24 A° in thickness are present in the light bands (Fig. 2). They are also found to be periodic acid-Schiff positive (Fig. 3).

scholarly journals ELECTRON MICROSCOPY OF THE UTERINE EPITHELIUM IN THE RABBIT

1962 ◽  
Vol 14 (1) ◽  
pp. 49-64 ◽  
Author(s):  
Jørgen Falck Larsen
Keyword(s):  
Electron Microscopy ◽  
Osmium Tetroxide ◽  
Periodic Acid ◽  
Hormonal Control ◽  
Uterine Epithelium ◽  
Chemical Effect ◽  
Periodic Acid Schiff ◽  
Multinucleated Cells ◽  
Hematoxylin And Eosin ◽  
In Pregnancy

The ultrastructure of the uterine epithelium has been studied in estrous, ovariectomized, pregnant, and pseudopregnant rabbits. Tissue for light microscopy was fixed in Bouin's solution and stained with hematoxylin and eosin, by the periodic acid-Schiff (PAS) method, and with methylene blue. Tissue for electron microscopy was fixed in 1 per cent osmium tetroxide in White's saline and embedded in Araldite. The uterine epithelium in estrus is comprised of ciliated and non-ciliated cells. After ovariectomy the epithelium becomes reduced in height and PAS-positive material disappears. Multinucleated cells are formed in the epithelium in pregnancy, pseudopregnancy, and in the non-pregnant horn in unilateral pregnancy. They degenerate during the 3rd week of pseudopregnancy and during the 4th week of pregnancy in the non-pregnant horn. The formation of multinucleated cells is believed to be under hormonal control. The uterine epithelium in contact with the blastocyst changes into a "symplasma," presumably under the influence of a local (chemical?) effect produced by the blastocyst. This change is not seen in pseudopregnancy nor in the non-pregnant horn in unilateral pregnancy. A complex infolding of the basal cell membrane of the epithelium accompanies the "symplasmic" change. The remaining uterine epithelium in pregnancy shows a well developed ergastoplasm suggesting a production of secretion materials, some of which may be available for absorption by the fetus through the yolk sac or paraplacental chorion.

scholarly journals HISTOCHEMICAL DEMONSTRATION OF SOME OXIDIZED MACROMOLECULES WITH THIOCARBOHYDRAZIDE (TCH) OR THIOSEMICARBAZIDE (TSC) AND OSMIUM TETROXIDE

1965 ◽  
Vol 13 (8) ◽  
pp. 629-639 ◽  
Author(s):  
ARNOLD M. SELIGMAN ◽  
JACOB S. HANKER ◽  
HANNAH WASSERKRUG ◽  
HERMINE DMOCHOWSKI ◽  
LIONEL KATZOFF
Keyword(s):  
Osmium Tetroxide ◽  
Periodic Acid ◽  
Acid Hydrazide ◽  
Histochemical Demonstration ◽  
Basement Membranes ◽  
Acid Oxidation ◽  
Cell Nuclei ◽  
Periodic Acid Schiff ◽  
Naphthoic Acid ◽  
Periodic Acid Oxidation

Thiocarbohydrazide (TCH) and thiosemicarbazide (TSC) have been introduced as osmiophilic reagents for demonstrating aldehyde-containing macromolecules, produced by periodic acid oxidation of tissue sections. Because of the remarkable properties of osmium black, excellent pigment qualities are provided for light microscopy and the advantages of an amorphous, nonvolatile, electron-opaque end product are available for electron microscopy. The present communication describes the method and gives the scope of the histochemical reaction. Thiocarbohydrazide is the more reactive of the two reagents and is capable of demonstrating a wider variety of oxidized macromolecules than does thiosemicarbazide. The periodic acid-thiocarbohydrazide-osmium tetroxide method (PATCO method) was compared with those using thiosemicarbazide (PATO method), periodic acid-Schiff (PAS method), and with the method involving 3-hydroxy-2-naphthoic acid hydrazide (NAH) followed by coupling with fast blue B (PA-NAH-FBB method). All four methods demonstrated certain active aldehydes equally well, such as those produced by periodic acid oxidation of glycogen, mucopolysaccharide of gut and mucoprotein of Descemet's membranes of the cornea. The stain with the PATO method was somewhat weaker than the other three methods for glycoprotein of the cuticular border of gut and of the brush border of kidney tubules, and colloid of the thyroid gland. The PATO method was even less effective in demonstrating mucoprotein of basement membranes in kidney and no reaction for collagen and reticulin was noted. The PATCO method was almost as effective with basement membranes in kidney as the PAS method, but was not equal to the PAS method in staining collagen and reticulin. Furthermore, the aldehyde produced on Feulgen hydrolysis of deoxyribonucleic acid (DNA) of cell nuclei was not demonstrable with either PATO or PATCO methods its contrast to positive results with the PAS and PA-NAH-FBB methods. This suggests that TSC and TCH have less reactive hydrazino groups than NAH. Results with the electron microscope will be published separately.

scholarly journals SPERMIOGENESIS IN CANCER CRABS

1969 ◽  
Vol 43 (3) ◽  
pp. 575-603 ◽  
Author(s):  
Susan G. Langreth
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Nucleic Acid ◽  
Sperm Cell ◽  
Periodic Acid ◽  
Light And Electron Microscopy ◽  
Mature Sperm ◽  
Periodic Acid Schiff ◽  
The Core ◽  
Inner Region

Spermiogenesis in Cancer crabs was studied by light and electron microscopy. The sperm are aflagellate, and when mature consist primarily of a spherical acrosome surrounded by the nucleus with its short radiating arms. The acrosome forms by a coalescence of periodic acid-Schiff-positive (PAS-positive) vesicles. During spermiogenesis one edge of the acrosomal vesicle invaginates to form a PAS-negative central core. The inner region of the acrosome bounding the core contains basic proteins which are not complexed to nucleic acid. The formation of an elaborate lattice-like complex of fused membranes, principally from membranes of the endoplasmic reticulum, is described. These membranes are later taken into the nucleus and subsequently degenerate. In late spermatids, when most of the cytoplasm is sloughed, the nuclear envelope and the cell membrane apparently fuse to become the limiting boundary over most of the sperm cell. In the mature sperm the chromatin of the nucleus and arms, which is Feulgen-positive, contains no detectable protein. The chromatin filaments appear clumped, branched, and anastomosed; morphologically, they resemble the DNA of bacterial nuclei. Mitochondria are absent or degenerate in mature sperm of Cancer crabs, but the centrioles persist in the nucleoplasm at the base of the acrosome.

scholarly journals Induction of pinocytosis in rat hepatocytes by partial hepatectomy.

1977 ◽  
Vol 72 (3) ◽  
pp. 695-706 ◽  
Author(s):  
M Mori ◽  
A B Novikoff
Keyword(s):  
Electron Microscopy ◽  
Body Weight ◽  
Partial Hepatectomy ◽  
Low Dose ◽  
Periodic Acid ◽  
Light And Electron Microscopy ◽  
Rat Hepatocytes ◽  
Lysosomal Hydrolases ◽  
Periodic Acid Schiff ◽  
Perfusion Fixation

Rat hepatocytes, normally not highly pinocytic cells, becomes so after partial hepatectomy when about two-thirds of the liver is removed. Droplets, up to 20 mum in diameter, develop, initially by addition to smaller pinocytic structures and later by fusion with lysosomes. The droplets contain a material with an electron microscope periodicity characteristic of fibrin; they are periodic acid Schiff-positive as is plasma. It is therefore reasonable to consider plasma glycoproteins to be major components of the droplets. The droplets are at all times membrane delimited, an observation possible only after perfusion fixation. The droplets are positive for three lysosomal hydrolases identified cytochemically: acid phosphatase, N-acetyl-beta-glucosaminidase, and beta-glucuronidase. From light and electron microscopy it is evident that these activities are acquired by fusion with lysosomes, mostly autophagic vacuoles and residual bodies both of which become very numerous after partial hepatectomy. Pinocytic structures are seen relatively infrequently in the hepatocytes of normal rats but a great many are present after partial hepatectomy. They are most easily observed if horseradish peroxidase (HRP) is intravenously injected before sacrifice and sections are incubated for HRP cytochemistry. The low dose of HRP employed (10 mg/100 g body weight) does not induce pinocytosis in controls, either untreated rats or rats subjected to laparotomy, including palpation of the liver. However, in partially hepatectomized rats even a much smaller dose of intravenous HRP (3.3 mg/100 g) visualizes the pinocytic structures in hepatocytes (coated vesicles, channels, cuplike bodies, and droplets). Kupffer cells pinocytose much HRP in both control and partially hepatectomized rats.

scholarly journals Phenotypic Modulation of Skeletal Muscle Fibers in LPIN1-Deficient Lipodystrophic (fld) Mice

2018 ◽  
Vol 56 (2) ◽  
pp. 322-331
Author(s):  
Rani S. Sellers ◽  
S. Radma Mahmood ◽  
Geoffrey S. Perumal ◽  
Frank P. Macaluso ◽  
Irwin J. Kurland
Keyword(s):  
Electron Microscopy ◽  
Skeletal Muscle ◽  
Fiber Type ◽  
Periodic Acid ◽  
Muscle Fibers ◽  
Myosin Atpase ◽  
Hybrid Fibers ◽  
Periodic Acid Schiff ◽  
Skeletal Muscle Fibers ◽  
Lipin 1

Lipin-1 ( Lpin1)–deficient lipodystrophic mice have scant and immature adipocytes and develop transient fatty liver early in life. Unlike normal mice, these mice cannot rely on stored triglycerides to generate adenosine triphosphate (ATP) from the β-oxidation of fatty acids during periods of fasting. To compensate, these mice store much higher amounts of glycogen in skeletal muscle and liver than wild-type mice in order to support energy needs during periods of fasting. Our studies demonstrated that there are phenotypic changes in skeletal muscle fibers that reflect an adaptation to this unique metabolic situation. The phenotype of skeletal muscle (soleus, gastrocnemius, plantaris, and extensor digitorum longus [EDL]) from Lpin1-/- was evaluated using various methods including immunohistochemistry for myosin heavy chains (Myh) 1, 2, 2a, 2b, and 2x; enzyme histochemistry for myosin ATPase, cytochrome-c oxidase (COX), and succinyl dehydrogenase (SDH); periodic acid–Schiff; and transmission electron microscopy. Fiber-type changes in the soleus muscle of Lpin1-/- mice were prominent and included decreased Myh1 expression with concomitant increases in Myh2 expression and myosin-ATPase activity; this change was associated with an increase in the presence of Myh1/2a or Myh1/2x hybrid fibers. Alterations in mitochondrial enzyme activity (COX and SDH) were apparent in the myofibers in the soleus, gastrocnemius, plantaris, and EDL muscles. Electron microscopy revealed increases in the subsarcolemmal mitochondrial mass in the muscles of Lpin1-/- mice. These data demonstrate that lipin-1 deficiency results in phenotypic fiber-specific modulation of skeletal muscle necessary for compensatory fuel utilization adaptations in lipodystrophy.

Mucopolysaccharides and the Calcification of the Scale of the Roach (Leuciscus rutilus)

1956 ◽  
Vol s3-97 (39) ◽  
pp. 329-332
Author(s):  
O. WALLIN
Keyword(s):  
Transition Zone ◽  
Great Increase ◽  
Periodic Acid ◽  
Mineral Matter ◽  
Large Crystal ◽  
Periodic Acid Schiff ◽  
Acid Mucopolysaccharides ◽  
Pas Reaction ◽  
Inorganic Fraction

1. The scale of the roach (Leuciscus rutilus L.) was examined for state of calcification, metachromasy, and reaction to the periodic acid / Schiff (PAS) test. 2. Metachromasy and a positive PAS-reaction imply acid mucopolysaccharides in the bony layer. 3. There is a great increase in these reactions and in reactions for bone salts in the transition zone between the uncalcified and calcified part of the bony layer. 4. These reactions imply that the bond between the osseoid and the inorganic fraction of the bony layer is through SO2- and PO4- groupings. 5. The fibrillary plate lacks metachromasy, but shows a positive PAS-reaction. Before calcification a strong orthochromasy points to acid groups in connexion with the collagen. Under the radii there is no orthochromasy, and the PAS-reaction is negative. 6. When calcifying, the fibrillary plate loses its orthochromasy and the mineral matter is deposited as large crystal-complexes. 7. In regenerating scales the reactions are weaker than in normal scales.

Sign in / Sign up

Export Citation Format

Share Document