Transient decondensation of chromatin in liver nuclei of rats treated with tannic acid

Bismuth (Bi) stains nucleoproteins (NPs) by interacting with available amino and primary phosphate groups. These two staining mechanisms are distinguishable by glutaraldehyde crosslinking (Fig. 1,2).Isolated mouse liver nuclei, extracted with salt and acid solutions, fixed in either formaldehyde (form.) or gl utaraldehyde (glut.) and stained with Bi, were viewed to determine the effect of the extractions on Bi stainina. Solubilized NPs were analyzed by SDS-polyacrylamide gel electrophoresis.Extraction with 0.14 M salt does not change the Bi staining characteristics (Fig. 3). 0.34 M salt reduces nucleolar (Nu) staining but has no effect on interchromatinic (IC) staining (Fig. 4). Proteins responsible for Nu and glut.- insensitive IC staining are removed when nuclei are extracted with 0.6 M salt (Fig. 5, 6). Low salt and acid extraction prevents Bi-Nu staining but has no effect on IC staining (Fig. 7). When nuclei are extracted with 0.6 M salt followed by low salt and acid, all Bi-staining components are removed (Fig. 8).

Download Full-text

A New Approach to the Demonstration of Oxidase-Localization Using Tannic Acid Or Catechin

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100073933 ◽

1973 ◽

Vol 31 ◽

pp. 698-699

Author(s):

V. Mizuhira ◽

Y. Futaesaku

Keyword(s):

Cross Section ◽

Tannic Acid ◽

Elastic Fiber ◽

The Other ◽

New Approach ◽

Tissue Block ◽

Active Reaction ◽

The Cross

Previously we reported that tannic acid is a very effective fixative for proteins including polypeptides. Especially, in the cross section of microtubules, thirteen submits in A-tubule and eleven in B-tubule could be observed very clearly. An elastic fiber could be demonstrated very clearly, as an electron opaque, homogeneous fiber. However, tannic acid did not penetrate into the deep portion of the tissue-block. So we tried Catechin. This shows almost the same chemical natures as that of proteins, as tannic acid. Moreover, we thought that catechin should have two active-reaction sites, one is phenol,and the other is catechole. Catechole site should react with osmium, to make Os- black. Phenol-site should react with peroxidase existing perhydroxide.

Download Full-text

Structural preservation and absolute contrast of catalase crystal sections prepared with tannic acid

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010007597x ◽

1983 ◽

Vol 41 ◽

pp. 448-449

Author(s):

C.W. Akey ◽

M. Szalay ◽

S.J. Edelstein

Keyword(s):

Tannic Acid ◽

Beef Heart ◽

X Rays ◽

Screw Axis ◽

General Applicability ◽

Thin Sections ◽

Beef Liver ◽

3 Dimensional ◽

Dimensional Reconstruction ◽

Structural Preservation

Three methods of obtaining 20 Å resolution in sectioned protein crystals have recently been described. They include tannic acid fixation, low temperature embedding and grid sectioning. To be useful for 3-dimensional reconstruction thin sections must possess suitable resolution, structural fidelity and a known contrast. Tannic acid fixation appears to satisfy the above criteria based on studies of crystals of Pseudomonas cytochrome oxidase, orthorhombic beef liver catalase and beef heart F1-ATPase. In order to develop methods with general applicability, we have concentrated our efforts on a trigonal modification of catalase which routinely demonstrated a resolution of 40 Å. The catalase system is particularly useful since a comparison with the structure recently solved with x-rays will permit evaluation of the accuracy of 3-D reconstructions of sectioned crystals.Initially, we re-evaluated the packing of trigonal catalase crystals studied by Longley. Images of the (001) plane are of particular interest since they give a projection down the 31-screw axis in space group P3121. Images obtained by the method of Longley or by tannic acid fixation are negatively contrasted since control experiments with orthorhombic catalase plates yield negatively stained specimens with conditions used for the larger trigonal crystals.

Download Full-text

Cell Fine Structure as Revealed in Dessicator-Dried Resinless Sections

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100099532 ◽

1981 ◽

Vol 39 ◽

pp. 622-623

Author(s):

R. P. Becker ◽

J. J. Wolosewick ◽

J. Ross-Stanton

Keyword(s):

Fine Structure ◽

Tannic Acid ◽

Three Dimensional ◽

Albino Rats ◽

Cell Organelles ◽

Vascular Perfusion ◽

Thin Sections ◽

Carbon Coated ◽

Embedding Medium ◽

Polyethylene Glycol 6000

Methodology has been introduced recently which allows transmission and scanning electron microscopy of cell fine structure in semi-thin sections unencumbered by an embedding medium. Images obtained from these “resinless” sections show a three-dimensional lattice of microtrabeculfee contiguous with cytoskeletal structures and membrane-bounded cell organelles. Visualization of these structures, especially of the matiiDra-nous components, can be facilitated by employing tannic acid in the fixation step and dessicator drying, as reported here.Albino rats were fixed by vascular perfusion with 2% glutaraldehyde or 1.5% depolymerized paraformaldehyde plus 2.5% glutaraldehyde in 0.1M sodium cacodylate (pH 7.4). Tissues were removed and minced in the fixative and stored overnight in fixative containing 4% tannic acid. The tissues were rinsed in buffer (0.2M cacodylate), exposed to 1% buffered osmium tetroxide, dehydrated in ethyl alcohol, and embedded in pure polyethylene glycol-6000 (PEG). Sections were cut on glass knives with a Sorvall MT-1 microtome and mounted onto poly-L-lysine, formvar-carbon coated grids while submerged in a solution of 95% ethanol containing 5% PEG.

Download Full-text

Extracellular filaments in the perineurium of the florida lobster, Panulirus argus

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100128237 ◽

1987 ◽

Vol 45 ◽

pp. 784-785

Author(s):

Roy J. Baerwald ◽

Lura C. Williamson

Keyword(s):

Blood Brain Barrier ◽

Glial Cells ◽

Tannic Acid ◽

Nerve Cord ◽

Ventral Nerve Cord ◽

Florida Keys ◽

Panulirus Argus ◽

Brain Barrier ◽

Cacodylate Buffer ◽

Nerve Cords

In arthropods the perineurium surrounds the neuropile, consists of modified glial cells, and is the morphological basis for the blood-brain barrier. The perineurium is surrounded by an acellular neural lamella, sometimes containing scattered collagen-like fibrils. This perineurial-neural lamellar complex is thought to occur ubiquitously throughout the arthropods. This report describes a SEM and TEM study of the sheath surrounding the ventral nerve cord of Panulirus argus.Juvenile P. argus were collected from the Florida Keys and maintained in marine aquaria. Nerve cords were fixed for TEM in Karnovsky's fixative and saturated tannic acid in 0.1 M Na-cacodylate buffer, pH = 7.4; post-fixed in 1.0% OsO4 in the same buffer; dehydrated through a graded series of ethanols; embedded in Epon-Araldite; and examined in a Philips 200 TEM. Nerve cords were fixed for SEM in a similar manner except that tannic acid was not used.

Download Full-text

Effects of Mg2+ on Chromatic Structure in Isolated Chicken Liver Nuclei

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100158194 ◽

1990 ◽

Vol 48 (3) ◽

pp. 130-131

Author(s):

Soichiro Arai ◽

Yuh H. Nakanishi

Keyword(s):

Chromatin Structure ◽

Room Temperature ◽

Osmium Tetroxide ◽

Divalent Cations ◽

Chromatin Condensation ◽

Chicken Liver ◽

Electron Microscopic ◽

Razor Blade ◽

Microscopic Studies ◽

Liver Nuclei

Although many electron microscopic studies on extracted chromatin have provided considerable information on chromatin condensation induced by divalent cations, there is only a little literature available on the effects of divalent cations on chromatin structure in intact nuclei. In the present study, the effects of Mg2+ on chromatin structure in isolated chicken liver nuclei were examined over a wide concentration range of Mg2+ by scanning electron microscopy.Nuclei were prepared from chicken liver by the method of Chauveau et al. with some modifications. The nuclei were suspended in 25 mM triethanolamine chloride buffer (pH7.4) with 1 mM EDTA or in the buffer with concentrations of MgCl2 varying from 1 to 50 mM. After incubation for 1 min at 0°C, glutaraldehyde was added to 1.8% and the nuclei were fixed for 1 h at 4°C. The fixed nuclei were mixed with 15% gelatin solution warmed at about 40°C, and kept at room temperature until the mixture set. The gelatin containing the nuclei was fixed with 2% glutaraldehyde for 2-4 h, and cut into small blocks. The gelatin blocks were conductive-stained with 2% tannic acid and 2% osmium tetroxide, dehydrated in a graded series of ethanol, and freeze-cracked with a razor blade in liquid nitrogen.

Download Full-text

Use of Factor VIII in the Treatment of Hemophilia

Thrombosis and Haemostasis ◽

10.1055/s-0038-1655471 ◽

1962 ◽

Vol 07 (02) ◽

pp. 230-238 ◽

Cited By ~ 1

Author(s):

A Pavlovsky ◽

H Peterson ◽

G Casillas ◽

C Simonetti ◽

A Martinez Canaveri ◽

...

Keyword(s):

Factor Viii ◽

Tannic Acid ◽

Deae Cellulose ◽

Fibrinolytic System ◽

Purification Method ◽

Fraction I

SummaryThis publication describes the results obtained by treatment of haemophiliacs with factor VIII preparations isolated from Cohn fraction I by use of tannic acid, FI-O-Ta.The authors stress the rapidity of the disappearance of factor VIII after injection. Transfusions are generally well tolerated. One reaction of the pyrogen type has been observed and also a case of activation of the fibrinolytic system.A second purification method by means of chromatography on DEAE-cellulose is described.

Download Full-text