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.

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).

Comparative strengths and weaknesses of peroxidase and colloidal gold in pre- and post-embedding immunolabeling techniques

1987 ◽  
Vol 45 ◽  
pp. 1002-1005
Author(s):  
D. R. Abrahamson ◽  
P. L. St.John ◽  
E. W. Perry
Keyword(s):  
Electron Microscopy ◽  
Horseradish Peroxidase ◽  
Light Microscopy ◽  
Colloidal Gold ◽  
Light Microscope ◽  
Ultrastructural Localization ◽  
The Other ◽  
Quantitative Studies ◽  
Dense Suspension ◽  
Antigen Localization

Antibodies coupled to tracers for electron microscopy have been instrumental in the ultrastructural localization of antigens within cells and tissues. Among the most popular tracers are horseradish peroxidase (HRP), an enzyme that yields an osmiophilic reaction product, and colloidal gold, an electron dense suspension of particles. Some advantages of IgG-HRP conjugates are that they are readily synthesized, relatively small, and the immunolabeling obtained in a given experiment can be evaluated in the light microscope. In contrast, colloidal gold conjugates are available in different size ranges and multiple labeling as well as quantitative studies can therefore be undertaken through particle counting. On the other hand, gold conjugates are generally larger than those of HRP but usually can not be visualized with light microscopy. Concern has been raised, however, that HRP reaction product, which is exquisitely sensitive when generated properly, may in some cases distribute to sites distant from the original binding of the conjugate and therefore result in spurious antigen localization.

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 Further Observations on the Fine Structure of Chrysochromulina Ericina Parke & Manton

1961 ◽  
Vol 41 (1) ◽  
pp. 145-155 ◽  
Author(s):  
I. Manton ◽  
G. F. Leedale
Keyword(s):  
Electron Microscopy ◽  
Fine Structure ◽  
Cell Surface ◽  
Light Microscopy ◽  
Flat Plate ◽  
Outer Layer ◽  
Living Cells ◽  
Layered Plates ◽  
Thin Sections ◽  
Micro Anatomy

C. ericina Parke & Manton has been re-investigated to add salient features of micro-anatomy from the electron microscopy of thin sections and also to add photographs of living cells taken with anoptral contrast light microscopy.The most important new observations concern the scales which are shown to be essentially two-layered plates in which the layers in the very large spined scales have become separated except at their edges, with the outer layer greatly hypertrophied to produce a hollow spine with a flared base closed at the bottom by a flat plate. The patterns of external marking on the two layers are very similar in both plate-scales and spines in this species and the orientation of both with respect to the cell surface has been demonstrated by a section of the scales in situ.

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.

Cytology and ultrastructure of Thanatephorus cucumeris and related taxa of the Rhizoctonia complex

1977 ◽  
Vol 55 (18) ◽  
pp. 2419-2436 ◽  
Author(s):  
C. C. Tu ◽  
James W. Kimbrough ◽  
H. C. Aldrich
Keyword(s):  
Electron Microscopy ◽  
Light Microscope ◽  
Light And Electron Microscopy ◽  
Middle Layer ◽  
Ultrastructural Level ◽  
Aniline Blue ◽  
Diploid Nucleus ◽  
Thanatephorus Cucumeris ◽  
Light Microscope Level ◽  
Endoplasmic Reticula

Cytological studies on the vegetative hyphae of members of the Rhizoctonia complex and basidial structures of Thanatephorus cucumeris were performed with light and electron microscopy. Vegetative cells of Thanatephorus and Waitea proved to be multinucleate, whereas those of Uthatobasidium, Ceratobasidium, Athelia. and Botryobasidium are binucleate.Dolipore septa of Thanatephorus, Waitea, Uthatobasidium, and Ceratobasidium are visible with the light microscope when stained with aniline blue in glycerine. Ultrastructurally, pore caps in these genera consisted of two-layered unit membranes, forming cisternae with an electron-dense middle layer. Dolipore septa of Athelia (S. rolfsii) and Botryobasidium are not visible in aniline blue at the light microscope level. At the ultrastructural level, there was an additional cisternal membrane making up a pore cap of three membranes. The fine structure of nuclei, mitochondria, endoplasmic reticula, vacuoles, and other organelles in the basidial structures of T. cucumeris was essentially the same as in other basidiomycetes.Karyogamy of two haploid nuclei occurs in the young basidia of T. cucumeris. The nuclear envelopes of both haploid nuclei break at their adjacent sides and fuse to form a diploid nucleus. After a short interphase, meiosis occurs. No leptotene was observed at prophase I, but a synaptinemal complex was evident and six pairs of chromosomes were observed throughout pachytene, diplotene, and diakinesis. The nuclear envelope disappears at metaphase I and a spindle appears. The second meiotic division is equational. Most of the mature and discharged spores are uninucleate.

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.

Demonstration of pulmonary vascular perfusion by electron and light microscopy

1993 ◽  
Vol 75 (4) ◽  
pp. 1877-1883 ◽  
Author(s):  
M. F. Konig ◽  
J. M. Lucocq ◽  
E. R. Weibel
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Light And Electron Microscopy ◽  
Rabbit Serum ◽  
Capillary Perfusion ◽  
Vascular Perfusion ◽  
Thin Sections ◽  
Gold Particles ◽  
Capillary Recruitment

To estimate the fraction of dense pulmonary capillary network that is perfused under physiological conditions, we developed a new method for the demonstration of in vivo capillary perfusion by light and electron microscopy. Blood plasma was labeled by 8-nm colloidal gold particles coated with rabbit serum albumin. In anesthetized rabbits, 4#x2013;5 ml of this tracer were injected into the right atrium. Two and 15 min later, the circulation was interrupted by a snare around the heart, and the lung was fixed by instillation with glutaraldehyde. Gold particles were found in the plasma space of alveolar capillaries as well as in other organs. A random sample of thin sections studied by electron microscopy revealed that the entire capillary bed of the lung was perfused at least with plasma within 2 min after tracer infusion. Light microscopy of silver-enhanced sections showed areas with different staining intensities but no obviously unperfused capillaries. The concept of capillary recruitment, which would require a significant fraction of capillaries unperfused at rest, may have to be reassessed to consider time factors as well as the two-phase nature of blood; red blood cells and plasma may take different paths.

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