scholarly journals THE LOCALIZATION OF ACID PHOSPHATASE IN RAT LIVER CELLS AS REVEALED BY COMBINED CYTOCHEMICAL STAINING AND ELECTRON MICROSCOPY

1961 ◽  
Vol 11 (1) ◽  
pp. 47-66 ◽  
Author(s):  
S. J. Holt ◽  
R. Marian Hicks
Keyword(s):  
Electron Microscopy ◽  
Acid Phosphatase ◽  
Light Microscope ◽  
Staining Pattern ◽  
Dense Body ◽  
Butyl Methacrylate ◽  
Nuclear Staining ◽  
Heterogeneous Distribution ◽  
Dense Bodies ◽  
Before And After

Discrete localization of stain in pericanalicular granules was found in 10 µ frozen sections of formol-phosphate-sucrose-fixed liver stained by the Gomori acid phosphatase technique and examined in the light microscope. The staining patterns, before and after treatment with Triton X-100 and lecithinase, were identical with those previously reported for formol-calcium-fixed material treated in the same way, and it can be assumed that the stained granules are identical with "lysosomes." Examination in the light microscope of the staining patterns and lead penetration in fixed blocks and slices of various dimensions showed nuclear staining and other artefacts to be present, produced by the different rates of penetration of the various components of the staining medium into the tissue. A uniform pericanalicular staining pattern could be obtained, however, with slices not more than 50 µ thick, into which the staining medium could penetrate rapidly from both faces. The staining pattern produced in 50 µ slices was the same both at pH 5.0 and pH 6.2, and was not altered by subsequent embedding of the stained material in butyl methacrylate. Electron microscopy showed the fine structure of fixed 50 µ frozen slices to be well preserved, but it deteriorated badly when they were incubated in the normal Gomori medium at pH 5.0 before postfixing in osmium tetroxide. After incubation in the Gomori medium at pH 6.2, the detailed morphology was substantially maintained. In both cases lead phosphate, the reaction product, was found in the pericanalicular regions of the cell, but only in the vacuolated dense bodies and never in the microbodies. Not every vacuolated dense body contained lead, and stained and unstained bodies were sometimes seen adjacent to each other. This heterogeneous distribution of stain within a morphologically homogeneous group of particles is consistent with de Duve's suggestion (9) that there is a heterogeneous distribution of enzymes within the lysosome population. It is concluded from these investigations that the vacuolated dense bodies seen in the electron microscope are the morphological counterparts of the "lysosomes" defined biochemically by de Duve.

The Time Relation Between The Haemostatic Aggregation Of Platelets And Their Release Reaction In Vivo

1981 ◽  
Author(s):  
P Görög ◽  
G V R Born
Keyword(s):  
Electron Microscopy ◽  
Dense Body ◽  
Feedback Mechanism ◽  
Mesenteric Arteries ◽  
Considerable Uncertainty ◽  
Release Reaction ◽  
Dense Bodies ◽  
Aggregation Of Platelets

The release reaction of platelets has been assumed to subserve a positive feedback mechanism responsible for their aggregation in haemostasis and thrombosis. This assumption is based mainly on in vitro experiments. Considerable uncertainty remains about the contribution of the release reaction to the initiation of haemostasis in vivo. The rapidity of the process and the presence of other tissues makes it impossible to follow the reaction quantitatively in vivo by methods which permit this in vitro. We have therefore applied quantitative electron microscopy to find out how quickly the concentration of dense bodies decreases in platelets during their haemostatic aggregation. In mice, platelets were enriched in dense bodies by pretreatment with serotonin.Mesenteric arteries were incised with a sharp blade. Bleeding was stopped by a micromanipulator-operated device about 15 sec and 60 sec after the cut. The cut segments were immediately fixed in situ with glutaraldehyde and postfixed. Serial sections were made for electron microscopy. Platelets isolated from peripheral blood of the same animal were prepared similarly. Electron micrographs were projected on to a television screen and numbers of dense bodies and total platelet areas were determined by an image analysing computer. After 15 sec there were no significant differences in numbers of dense bodies in platelets from different parts of the haemostatic plugs (8.31±0.57/100 μ2 mean ± s.e.m.) and in platelets from the blood (8.93±0.38). On the other hand, after 60 sec the parts furthest from the cut contained fewer dense bodies than the nearer parts and the overall dense body number (5.86±0.05) was considerably smaller (p<0.001) than that of platelets from the blood (14.45±0.09).The results suggest that haemostatic aggregation of platelets does not initially depend on their release reaction.

scholarly journals QUANTITATIVE CHARACTERIZATION OF DENSE BODY, AUTOPHAGIC VACUOLE, AND ACID PHOSPHATASE-BEARING PARTICLE POPULATIONS DURING THE EARLY PHASES OF GLUCAGON-INDUCED AUTOPHAGY IN RAT LIVER

1971 ◽  
Vol 48 (3) ◽  
pp. 473-489 ◽  
Author(s):  
Russell L. Deter
Keyword(s):  
Acid Phosphatase ◽  
Dense Body ◽  
Autophagic Vacuole ◽  
Type I ◽  
Type Ii ◽  
Average Volume ◽  
Quantitative Characterization ◽  
Autophagic Vacuoles ◽  
Dense Bodies

Quantitative characterization of dense body, autophagic vacuole and acid phosphatase-bearing particle populations of rat liver have been made at 10 min intervals during the first 50 min following the intraperitoneal administration of glucagon. Beginning 10 to 20 min postinjection, increases in the number of autophagic vacuoles and in the osmotic sensitivity of acid phosphatase-bearing particles were observed, associated with a progressive disappearance of dense bodies. These changes appeared to reach a maximum 50 min after treatment. The average volume of autophagic vacuoles was found to be 440–870% greater than that of normal dense bodies during this time period. No consistent change in total acid phosphatase activity was noted. A detailed study of autophagic vacuole profile populations revealed the presence of five different types of profiles, two of which, types I and II, accounted for 76.3–94.4% of the profiles examined. Type I profiles primarily contained elements of the endoplasmic reticulum, free ribosomes, and ground cytoplasm. Type II profiles had mitochondrial profiles as their principal constituent, but endoplasmic reticulum and free ribosomes were also seen. At all time points type I profiles predominated, comprising 55–69% of the profiles found. Both profile types were bounded by single and double limiting membranes, the former being predominate. A time-dependent change in the ratio of single to double membrane-limited profiles could not be demonstrated. Morphometric parameters derived from profile size distributions indicated that the number of types I and II autophagic vacuoles increased with time, the rate being greater for the type II particle, except between 40 and 50 min. The average volume of the type II autophagic vacuole was consistently greater than that of the type I.

scholarly journals FINE STRUCTURAL LOCALIZATION OF ACID AND ALKALINE PHOSPHATASES IN CELLS OF RABBIT BLOOD AND BONE MARROW

1967 ◽  
Vol 15 (6) ◽  
pp. 311-334 ◽  
Author(s):  
B. K. WETZEL ◽  
S. S. SPICER ◽  
R. G. HORN
Keyword(s):  
Alkaline Phosphatase ◽  
Acid Phosphatase ◽  
Phosphatase Activity ◽  
Acid Phosphatase Activity ◽  
Mononuclear Cells ◽  
Nuclear Staining ◽  
Alkaline Phosphatases ◽  
Structural Localization ◽  
Dense Granules ◽  
Dense Bodies

In rabbit heterophils, acid phosphatase activity occurs in primary (azurophil) granules which predominate in early cells and persist in mature cells and in tertiary granules which are seen only in mature cells. Alkaline phosphatase activity occurs in secondary granules which appear in intermediate heterophils and later predominate in mature cells. Acid phosphatase activity in heterophil Golgi zones coincides developmentally with the genesis of primary and, later, tertiary granules, whereas alkaline phosphatase in the Golgi complex coincides with secondary granulogenesis. In developing eosinophils, acid phosphatase reaction product occurs in Golgi elements, rims the spherical precursors of angular, mature granules and appears inconsistently within mature granules. Basophil myelocytes show acid phosphatase in Golgi elements but not in specific granules. Additional acid phosphatase reactive structures include: granules of mononuclear cells; phagocytic vacuoles in macrophages; autophagic vacuoles in maturing erythroid cells; small dense granules of platelets; dense bodies in lipocytes; and Golgi elements of mononuclear cells, macrophages, nucleated red cells, megakaryocytes and lipocytes. Localized deposits were absent in control specimens except for enzyme-independent nuclear staining in alkaline phosphatase preparations.

scholarly journals ELECTRON MICROSCOPY OF LYSOSOME-RICH FRACTIONS FROM RAT THYMUS ISOLATED BY DENSITY-GRADIENT CENTRIFUGATION BEFORE AND AFTER WHOLE-BODY X-IRRADIATION

1962 ◽  
Vol 13 (2) ◽  
pp. 253-260 ◽  
Author(s):  
Y. E. Rahman
Keyword(s):  
Electron Microscopy ◽  
Acid Phosphatase ◽  
Cytochrome Oxidase ◽  
Density Gradient ◽  
Density Gradient Centrifugation ◽  
Gradient Centrifugation ◽  
Whole Body ◽  
Rat Tissues ◽  
Before And After ◽  
X Irradiation

Fractions from rat thymuses were isolated by sucrose density-gradient centrifugation, before and after 1000 r whole-body x-irradiation, and examined by electron microscopy. Cytochrome oxidase and acid phosphatase activities of these fractions were tested as well. Electron-opaque bodies with diameters ranging from 0.10 to 0.35 µ, with a mean of 0.25 µ, were found in fractions having high acid phosphatase activity, while the fractions rich in cytochrome oxidase consisted mostly of mitochondria. After irradiation, there was an increased ratio of dense bodies to mitochondria. These particles are considered to be lysosomes similar to those identified in other rat tissues. Their relationship to the mitochondria is discussed.

scholarly journals DEMONSTRATION OF ACID PHOSPHATASE-CONTAINING GRANULES AND CYTOPLASMIC BODIES IN THE EPITHELIUM OF FOETAL RAT DUODENUM DURING CERTAIN STAGES OF DIFFERENTIATION

1963 ◽  
Vol 18 (2) ◽  
pp. 251-265 ◽  
Author(s):  
O. Behnke
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Acid Phosphatase ◽  
Parallel Study ◽  
Frozen Sections ◽  
Cytoplasmic Bodies ◽  
Dense Bodies ◽  
Foetal Rat ◽  
Rat Duodenum ◽  
Myelin Figures

Dense cytoplasmic bodies surrounded by one or two unit membranes and containing mitochondria, vesicles, ribosomes, rough and smooth surfaced endoplasmic reticulum, and lamellated membranes (myelin figures) have been observed in the differentiating mucosa of the duodenum of rat foetuses by electron microscopy. Generally, the cytoplasmic components in the bodies seem to be in varying stages of disintegration. The bodies are found in greatest number on the 17th and 18th day of gestation, i.e. at the onset of differentiation. At this period of development the epithelium is stratified, and the villus formation is initiated by invagination of the epithelium by buds of mesenchyme followed by a splitting of the epithelium along the sides of the invaginations. When the villi have formed, the stratified epithelium has changed to the simple columnar type and the dense bodies have largely disappeared. Simultaneously, the lumen has widened considerably. In a parallel study with the light microscope, frozen sections incubated for the demonstration of acid phosphatase activity revealed the reaction product to be localized in bodies of the same size and distribution as the dense bodies found by electron microscopy. Hence, it seems that the bodies are altered and enlarged lysosomes (cytolysomes) active during the intensive differentiative events in the small intestine during the last part of intra-uterine life.

scholarly journals COMBINED CYTOCHEMICAL AND ELECTRON MICROSCOPIC DEMONSTRATION OF ß-GLUCURONIDASE ACTIVITY IN RAT LIVER WITH THE USE OF A SIMULTANEOUS COUPLING AZO DYE TECHNIQUE

1968 ◽  
Vol 36 (2) ◽  
pp. 289-297 ◽  
Author(s):  
Masando Hayashi ◽  
Tsuranobu Shirahama ◽  
Alan S. Cohen
Keyword(s):  
Electron Microscopy ◽  
Rat Liver ◽  
Dense Body ◽  
Electron Microscopic ◽  
Glucuronidase Activity ◽  
Dense Bodies ◽  
The Matrix ◽  
Matrix Control ◽  
Parenchymal Cells ◽  
Cytochemical Technique

Rat liver was fixed in formal-cacodylate-sucrose and frozen sections were incubated in a simultaneous-coupling medium containing naphthol AS-BI glucuronide as substrate and hexazonium pararosanilin as the diazo reagent. By light microscopy, the sections demonstrated ß-glucuronidase activity as red discrete granules in the pericanalicular cytoplasm and as a generalized cytoplasmic stain in the parenchymal cells. Brief treatment of sections in cold ethanol prior to incubation markedly enhanced the staining for the enzyme and made it possible to demonstrate sufficient amounts of the reaction product in sections embedded in epoxy resin following dehydration and propylene oxide treatment. Electron microscopy revealed that the reaction product was moderately electron opaque and deposited in greater amounts in the vacuolated dense bodies and occasionally in the dense bodies which did not show obvious vacuoles. In each dense body, the deposits occurred preferentially at the edge as well as in the area surrounding the vacuoles in the matrix. Control sections incubated in the presence of glucosaccharo-1:4-lactone were devoid of the reaction product. No deposits of the reaction product were found in the nucleus, mitochondria, or microbodies. The limitations of the present cytochemical technique for use in electron microscopy are briefly discussed.

scholarly journals OBSERVATIONS ON THE FINE STRUCTURE OF PHARYNGEAL MUSCLE IN THE PLANARIAN DUGESIA TIGRINA

1963 ◽  
Vol 18 (3) ◽  
pp. 651-662 ◽  
Author(s):  
Edith Krugelis MacRae
Keyword(s):  
Electron Microscopy ◽  
Plasma Membrane ◽  
Fine Structure ◽  
Osmium Tetroxide ◽  
Dense Body ◽  
Longitudinal Axis ◽  
Pharyngeal Muscle ◽  
Dugesia Tigrina ◽  
Dense Bodies ◽  
And Function

Pharyngeal muscle of the planarian Dugesia tigrina was studied by electron microscopy after osmium tetroxide fixation. The muscle cell was observed to contain one myofibril or bundle of myofilaments parallel to its longitudinal axis. The myofilaments were of two types, different in size and distribution. No Z lines or myofilament organization into cross or helical striations were seen. Dense bodies were seen as projections from an invagination of the plasma membrane and as dense lines parallel to the myofilaments. The muscle cells are surrounded by a plasma membrane which is structurally associated with dense body projections, with vesicles and cisternae of sarcoplasmic reticulum, and with synaptic nerve endings. The cell has sarcoplasmic projections perpendicular to its long axis; these projections are seen to contain the nucleus or mitochondria and granules. Mitochondria and granules are also seen in a sarcoplasm rim around the fibril. The dense bodies may serve as attachment for thin myofilaments and function in transmission of stimuli from plasma membrane to the interior of the fibril.

The Precipitation of Si in AlCuSi Thin Films

1973 ◽  
Vol 31 ◽  
pp. 34-35 ◽  
Author(s):  
R. M. Anderson
Keyword(s):  
Electron Microscopy ◽  
Thin Films ◽  
Heat Treatment ◽  
Solid Solution ◽  
Integrated Circuit ◽  
Copper Film ◽  
Solution Concentration ◽  
X Ray ◽  
Silicon Thin Films ◽  
Before And After

Aluminum-copper-silicon thin films have been considered as an interconnection metallurgy for integrated circuit applications. Various schemes have been proposed to incorporate small percent-ages of silicon into films that typically contain two to five percent copper. We undertook a study of the total effect of silicon on the aluminum copper film as revealed by transmission electron microscopy, scanning electron microscopy, x-ray diffraction and ion microprobe techniques as a function of the various deposition methods.X-ray investigations noted a change in solid solution concentration as a function of Si content before and after heat-treatment. The amount of solid solution in the Al increased with heat-treatment for films with ≥2% silicon and decreased for films <2% silicon.

Copper Localization in Cirrhotic Rat Liver by Scanning Electron Microscopy

1969 ◽  
Vol 27 ◽  
pp. 38-39 ◽  
Author(s):  
J. C. Russ ◽  
E. McNatt
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Electron Microscope ◽  
Copper Acetate ◽  
Lead Citrate ◽  
Dense Bodies ◽  
Transmission Electron ◽  
Electron Mode ◽  
Scanning Electron

In order to study the retention of copper in cirrhotic liver, rats were made cirrhotic by carbon tetrachloride inhalation twice weekly for three months and fed 0.2% copper acetate ad libidum in drinking water for one month. The liver tissue was fixed in osmium, sectioned approximately 2000 Å thick, and stained with lead citrate. The section was examined in a scanning electron microscope (JEOLCO JSM-2) in the transmission electron mode.Figure 1 shows a typical area that includes a red blood cell in a sinusoid, a disse, and a portion of the cytoplasm of a hepatocyte which contains several mitochondria, peribiliary dense bodies, glycogen granules, and endoplasmic reticulum.

Epoxy Resin Embedment

1968 ◽  
Vol 26 ◽  
pp. 100-101
Author(s):  
J. G. Adams ◽  
M. M. Campbell ◽  
H. Thomas ◽  
J. J. Ghldonl
Keyword(s):  
Electron Microscopy ◽  
Electron Beam ◽  
Epoxy Resin ◽  
Light Microscope ◽  
Epoxy Resins ◽  
Image Contrast ◽  
Tissue Preservation ◽  
Resin System ◽  
Excellent Quality

Since the introduction of epoxy resins as embedding material for electron microscopy, the list of new formulations and variations of widely accepted mixtures has grown rapidly. Described here is a resin system utilizing Maraglas 655, Dow D.E.R. 732, DDSA, and BDMA, which is a variation of the mixtures of Lockwood and Erlandson. In the development of the mixture, the Maraglas and the Dow resins were tested in 3 different volumetric proportions, 6:4, 7:3, and 8:2. Cutting qualities and characteristics of stability in the electron beam and image contrast were evaluated for these epoxy mixtures with anhydride (DDSA) to epoxy ratios of 0.4, 0.55, and 0.7. Each mixture was polymerized overnight at 60°C with 2% and 3% BDMA.Although the differences among the test resins were slight in terms of cutting ease, general tissue preservation, and stability in the beam, the 7:3 Maraglas to D.E.R. 732 ratio at an anhydride to epoxy ratio of 0.55 polymerized with 3% BDMA proved to be most consistent. The resulting plastic is relatively hard and somewhat brittle which necessitates trimming and facing the block slowly and cautiously to avoid chipping. Sections up to about 2 microns in thickness can be cut and stained with any of several light microscope stains and excellent quality light photomicrographs can be taken of such sections (Fig. 1).

Sign in / Sign up

Export Citation Format

Share Document