scholarly journals Simultaneous localization of proteoglycan by light and electron microscopy using toluidine blue O. A study of epiphyseal cartilage.

1976 ◽  
Vol 24 (5) ◽  
pp. 621-629 ◽  
Author(s):  
N Shepard ◽  
N Mitchell
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Toluidine Blue ◽  
Light And Electron Microscopy ◽  
Uranyl Acetate ◽  
Collagen Fibrils ◽  
Epiphyseal Cartilage ◽  
Fixation Method ◽  
Fixed Tissue

The simultaneous localization of proteoglycan by light and electron microscopy was demonstrated by fixing epiphyseal cartilage in a glutaraldehyde toluidine blue O solution. Sections cut for light microscopy viewing and those cut for electron microscopy required no further staining, although, in the latter case, staining with uranyl acetate and lead improved the overall contrast. By this technique, electron-dense structures were seen concentrated about the cells which were actively synthesizing matrix, and these structures appeared to bind collagen fibrils. Similar structures were not seen in conventionally fixed tissue. They could also not be identified when the specimens were previously incubated with the proteoglycan-digesting enzyme, papain, prior to toluidine blue O fixation. The toluidine blue O fixation method, unlike conventional fixation and staining, retained proteoglycan in the pericellular areas of actively synthesizing cells and made it visible by light and electron microscopy. It appears that proteoglycans is both precipitated and stained by the presence of toluidine blue O during fixation.

Ultrastructure of Angiosarcoma of Mouse Tail

1980 ◽  
Vol 38 ◽  
pp. 740-741
Author(s):  
White Yvonne ◽  
Winslow Sheldon ◽  
James W. Townsend ◽  
Neil A. Littlefield
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Light Microscopy ◽  
Toluidine Blue ◽  
Female Mouse ◽  
Osmium Tetroxide ◽  
Uranyl Acetate ◽  
Lead Citrate ◽  
Ultrathin Sections ◽  
Resin Mixture

Spontaneous neoplasms rarely occur on the tails of BALB/cStCrlfC3H/Nctr mice, but the neoplasm most frequently observed is a locally invasive non-metastatic angiosarcoma. In this case a female mouse weighing 33.2 g, 706 days of age, presented a soft, red, irregularly-shaped mass, measuring 18 mm in its greatest dimension, in the subcutis of the base of the tail. A portion of the tail tumor was taken for electron microscopy and the remainder was processed for light microscopy. The tissue processed for electron microscopy was fixed in 4% cacodyl ate-buffered glutaraldehyde, post-fixed in 1% osmium tetroxide, dehydrated in a graded series of ethanol solutions, and embedded in Epon-Araldite resin mixture. Sections of 1 μm were stained with toluidine blue for light microscopy and ultrathin sections of 100 nm were stained with uranyl acetate and lead citrate, then examined with a Philips EM201 electron microscope .

scholarly journals Dense HRP filling in pre-fixed brain tissue for light and electron microscopy.

1987 ◽  
Vol 35 (10) ◽  
pp. 1127-1136 ◽  
Author(s):  
G H Kageyama ◽  
R L Meyer
Keyword(s):  
Electron Microscopy ◽  
Optic Nerve ◽  
High Resolution ◽  
Light Microscopy ◽  
Glial Cells ◽  
Light And Electron Microscopy ◽  
Polyethylene Tube ◽  
Fixed Tissue ◽  
Cat Brain

The use of neuroanatomical markers in tissues that have been pre-fixed has been virtually ignored, even though this approach could offer certain advantages over in vivo methods, in terms of convenience of application and choice of markers. We have found that HRP can be used on well-fixed brains of cats and goldfish to fill neurons, dendrites, axons, terminals, glial cells, and glial processes for high-resolution light microscopy and electron microscopy. Best results were obtained using brains that were perfusion-fixed with 2.5% depolymerized paraformaldehyde and 1.5% glutaraldehyde. Two methods of HRP application were used: optically guided injections of microliter quantities into various regions of cat brain, and optic nerve fills in goldfish by attaching an HRP-filled polyethylene tube for periods of 1 day to 2 weeks. HRP applied in these ways to pre-fixed tissue was found to fill neurons or glial cells with solid label in the anterograde and retrograde directions.

Effects of EDTA, Hyaluronidase and Collagenase on Epiphyseal Cartilage Matrix

1968 ◽  
Vol 26 ◽  
pp. 56-57
Author(s):  
H. Clarke Anderson ◽  
Priscilla R. Coulter
Keyword(s):  
Light And Electron Microscopy ◽  
Matrix Vesicles ◽  
Cartilage Matrix ◽  
Collagen Fibrils ◽  
Epiphyseal Cartilage ◽  
Dense Matrix ◽  
Type Iv ◽  
Bovine Testicular Hyaluronidase ◽  
The Matrix ◽  
Testicular Hyaluronidase

Epiphyseal cartilage matrix contains fibrils and particles of at least 5 different types: 1. Banded collagen fibrils, present throughout the matrix, but not seen in the lacunae. 2. Non-periodic fine fibrils <100Å in diameter (Fig. 1), which are most notable in the lacunae, and may represent immature collagen. 3. Electron dense matrix granules (Fig. 1) which are often attached to fine fibrils and collagen fibrils, and probably contain protein-polysaccharide although the possibility of a mineral content has not been excluded. 4. Matrix vesicles (Fig. 2) which show a selective distribution throughout the epiphysis, and may play a role in calcification. 5. Needle-like apatite crystals (Fig. 2).Blocks of formalin-fixed epiphysis from weanling mice were digested with the following agents in 0.1M phosphate buffer: a) 5% ethylenediaminetetraacetate (EDTA) at pH 8.3, b) 0.015% bovine testicular hyaluronidase (Sigma, type IV, 750 units/mg) at pH 5.5, and c) 0.1% collagenase (Worthington, chromatograhically pure, 200 units/mg) at pH 7.4. All digestions were carried out at 37°C overnight. Following digestion tissues were examined by light and electron microscopy to determine changes in the various fibrils and particles of the matrix.

The Response of Testis Cells to Low Dose X-Irradiation

1981 ◽  
Vol 39 ◽  
pp. 542-543
Author(s):  
D. E. Philpott ◽  
W. Sapp ◽  
C. Williams ◽  
Joann Stevenson ◽  
S. Black
Keyword(s):  
Electron Microscopy ◽  
Stem Cell ◽  
Light Microscopy ◽  
Dose Level ◽  
Cross Sections ◽  
Low Dose ◽  
Light And Electron Microscopy ◽  
Cellular Responses ◽  
Post Irradiation ◽  
X Irradiation

The response of spermatogonial cells to X-irradiation is well documented. It has been shown that there is a radiation resistent stem cell (As) which, after irradiation, replenishes the seminiferous epithelium. Most investigations in this area have dealt with radiation dosages of 100R or more. This study was undertaken to observe cellular responses at doses less than 100R of X-irradiation utilizing a system in which the tissue can be used for light and electron microscopy.Brown B6D2F1 mice aged 16 weeks were exposed to X-irradiation (225KeV; 15mA; filter 0.35 Cu; 50-60 R/min). Four mice were irradiated at each dose level between 1 and 100 rads. Testes were removed 3 days post-irradiation, fixed, and embedded. Sections were cut at 2 microns for light microscopy. After staining, surviving spermatogonia were identified and counted in tubule cross sections. The surviving fraction of spermatogonia compared to control, S/S0, was plotted against dose to give the curve shown in Fig. 1.

Solving the mechanisms responsible for organelle behavior in vivo by same-cell correlative light and electron microscopy

1992 ◽  
Vol 50 (1) ◽  
pp. 14-15
Author(s):  
Conly L. Rieder
Keyword(s):  
Electron Microscopy ◽  
Quantitative Analysis ◽  
Light Microscopy ◽  
Living Cell ◽  
Light And Electron Microscopy ◽  
Time Behavior ◽  
Dynamic Interactions ◽  
Positive Identification ◽  
Cellular Components

The behavior of many cellular components, and their dynamic interactions, can be characterized in the living cell with considerable spatial and temporal resolution by video-enhanced light microscopy (video-LM). Indeed, under the appropriate conditions video-LM can be used to determine the real-time behavior of organelles ≤ 25-nm in diameter (e.g., individual microtubules—see). However, when pushed to its limit the structures and components observed within the cell by video-LM cannot be resolved nor necessarily even identified, only detected. Positive identification and a quantitative analysis often requires the corresponding electron microcopy (EM).

Development of an in vitro Model for the Study of the Response of Equine Tendon Fibroblasts to Injury and Medication

1997 ◽  
Vol 10 (01) ◽  
pp. 6-11 ◽  
Author(s):  
R. F. Rosenbusch ◽  
L. C. Booth ◽  
L. A. Dahlgren
Keyword(s):  
Electron Microscopy ◽  
Extracellular Matrix ◽  
Light Microscopy ◽  
Light And Electron Microscopy ◽  
Tendon Healing ◽  
Matrix Proteins ◽  
Isolated Cells ◽  
Superficial Digital Flexor Tendon ◽  
Vitro Model

SummaryEquine tendon fibroblasts were isolated from explants of superficial digital flexor tendon, subcultured and maintained in monolayers. The cells were characterized by light microscopy, electron microscopy and radiolabel studies for proteoglycan production. Two predominant cell morphologies were identified. The cells dedifferentiated toward a more spindle shape with repeated subcultures. Equine tendon fibroblasts were successfully cryopreserved and subsequently subcultured. The ability to produce proteoglycan was preserved.The isolated cells were identified as fibroblasts, based on their characteristic shape by light microscopy and ultrastructure and the active production of extracellular matrix proteins. Abundant rough endoplasmic reticulum and the production of extracellular matrix products demonstrated active protein production and export. Proteoglycans were measurable via liquid scintillation counting in both the cell-associated fraction and free in the supernatant. This model is currently being utilized to study the effects of polysulfated glycosaminoglycan on tendon healing. Future uses include studying the effects of other pharmaceuticals, such as hyaluronic acid, on tendon healing.A model was developed for in vitro investigations into tendon healing. Fibroblasts were isolated from equine superficial digital flexor tendons and maintained in monolayer culture. The tenocytes were characterized via light and electron microscopy. Proteoglycan production was measured, using radio-label techniques. The fibroblasts were cryopreserved and subsequently subcultured. The cells maintained their capacity for proteoglycan production, following repeated subculturing and cryopreservation.

scholarly journals An Easy Path for Correlative Electron and Super-Resolution Light Microscopy

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Dorothea Pinotsi ◽  
Simona Rodighiero ◽  
Silvia Campioni ◽  
Gabor Csucs
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Light Microscopy ◽  
Amyloid Fibrils ◽  
Light And Electron Microscopy ◽  
Super Resolution ◽  
Wide Field ◽  
Set Up ◽  
Bio Compatibility ◽  
Scanning Electron

Abstract A number of new Correlative Light and Electron Microscopy approaches have been developed over the past years, offering the opportunity to combine the specificity and bio-compatibility of light microscopy with the high resolution achieved in electron microscopy. More recently, these approaches have taken one step further and also super-resolution light microscopy was combined with transmission or scanning electron microscopy. This combination usually requires moving the specimen between different imaging systems, an expensive set-up and relatively complicated imaging workflows. Here we present a way to overcome these difficulties by exploiting a commercially available wide-field fluorescence microscope integrated in the specimen chamber of a Scanning Electron Microscope (SEM) to perform correlative LM/EM studies. Super-resolution light microscopy was achieved by using a recently developed algorithm - the Super-Resolution Radial Fluctuations (SRRF) - to improve the resolution of diffraction limited fluorescent images. With this combination of hardware/software it is possible to obtain correlative super-resolution light and scanning electron microscopy images in an easy and fast way. The imaging workflow is described and demonstrated on fluorescently labelled amyloid fibrils, fibrillar protein aggregates linked to the onset of multiple neurodegenerative diseases, revealing information about their polymorphism.

scholarly journals THE GLOMERULUS IN EXPERIMENTAL RENAL DISEASE IN RATS AS OBSERVED BY LIGHT AND ELECTRON MICROSCOPY

1955 ◽  
Vol 102 (5) ◽  
pp. 573-580 ◽  
Author(s):  
Carolyn F. Piel ◽  
Luther Dong ◽  
F.W.S. Modern ◽  
Joseph R. Goodman ◽  
Roger Moore
Keyword(s):  
Electron Microscopy ◽  
Endothelial Cells ◽  
Basement Membrane ◽  
Light Microscopy ◽  
Light And Electron Microscopy ◽  
Electron Microscopic ◽  
Crescent Formation ◽  
Colloidal Iron ◽  
Narrow Channels ◽  
Glomerular Capillary

Nephrotoxic serum disease in rats has been studied by light and electron microscopy from 1 hour to 10 weeks after production of the disease. By light microscopy leucocytic infiltration of the glomerular capillary was observed between the 3rd and 6th hour. At 6 hours an increase in colloidal iron-positive material was observed coating the extraluminal surface of the capillaries. Also at this time swelling of the endothelial cells becomes prominent. By 72 hours, thickening of the basement membrane was observed. Glomerular capillary thrombi were observed in approximately half the tissue examined in the first 2 weeks of disease. 50 per cent of the animals showed severe chronic lesions, exudation into the capsular space, crescent formation, and obliteration of glomeruli. At 1 hour electron microscopic pictures showed that osmophilic material may line the foot processes of the epithelial cells and obliterate all but narrow channels of the space between the feet. By 6 hours thickening of the basement membrane was prominent. This change persisted throughout 10 weeks of observation. The tissue from animals which had severe chronic alterations by light microscopy revealed changes which could not be interpreted at this time.

scholarly journals A tribute to Shinya Inoue and innovation in light microscopy

2004 ◽  
Vol 165 (1) ◽  
pp. 21-26 ◽  
Author(s):  
Karen R. Dell ◽  
Ronald D. Vale
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Light Microscope ◽  
Light And Electron Microscopy ◽  
Single Molecules ◽  
Living Cells ◽  
Dynamic Processes ◽  
New Techniques

The 2003 International Prize for Biology was awarded to Shinya Inoue for his pioneering work in visualizing dynamic processes within living cells using the light microscope. He and his scientific descendants are now pushing light microscopy even further by developing new techniques such as imaging single molecules, visualizing processes in living animals, and correlating results from light and electron microscopy.

Scale structure and taxonomy of some species of Raphidocystis, Raphidiophrys, and Pompholyxophrys (Heliozoea) including descriptions of six new taxa

1985 ◽  
Vol 63 (8) ◽  
pp. 1944-1961 ◽  
Author(s):  
K. H. Nicholls ◽  
M. Dürrschmidt
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
New Zealand ◽  
Light Microscopy ◽  
Light And Electron Microscopy ◽  
Scale Structure ◽  
Taxonomic Descriptions ◽  
Transmission Electron ◽  
New Information ◽  
Freshwater Habitats

Sixteen taxa of the genera Raphidocystis, Raphidiophrys, and Pompholyxophrys from freshwater habitats in Canada, Chile, and New Zealand were studied by light and electron microscopy. Six taxa are described as new: Raphidocystis glabra, Raphidiophrys minuta, Raphidiophrys orbicularis ssp. orbicularis, R. orbicularis ssp. ovalis, Pompholyxophrys stellata, and P. ossea. New information on scale structure and arrangement based on scanning and transmission electron microscopy amplifies the taxonomic descriptions of Raphidiophrys ambigua, R. pallida, R. elegans, R. intermedia, R. marginata, R. symmetrica, Pompholyxophrys punicea, P. exigua, and P. ovuligera, which were previously imperfectly known by light microscopy only.

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