scholarly journals STRUCTURE IN NUCLEATED ERYTHROCYTES

1961 ◽  
Vol 9 (3) ◽  
pp. 671-687 ◽  
Author(s):  
Howard G. Davies
Keyword(s):  
Electron Microscopy ◽  
Nuclear Envelope ◽  
Light Microscope ◽  
Phase Contrast ◽  
Somatic Cells ◽  
Chemical Substance ◽  
Feulgen Staining ◽  
Nuclear Pores ◽  
Specific Absorption ◽  
Static Pictures

The structure of the nucleated erythrocyte of frog and chicken has been investigated by electron microscopy and correlated with the distribution of haemoglobin and DNA-containing material determined by haem absorption and Feulgen staining in the light microscope. The nuclei of both species are found to contain haemoglobin which is continuous with the haemoglobin in the cytoplasm through holes or pores in the nuclear envelope. In addition the nucleus of the frog erythrocyte sometimes contains a single invagination which is lined by the nuclear envelope. The structure of the nuclear envelope and the pores and the organisation of the nucleus are similar to those already described for other somatic cells. Erythrocytes differ from the cells previously studied in that a continuity, via the nuclear pores, of chemical substance in the interior of the nucleus and in the cytoplasm can be directly demonstrated. This is due to the fact that the cytoplasm of erythrocytes is simple, consisting predominantly of haemoglobin, and that haemoglobin is easily recognised by its specific absorption. The static pictures obtained by electron microscopy have been supplemented by observations in phase-contrast of the changes in refraction of the cell contents due to the diffusion of the haemoglobin from the nucleus into the cytoplasm during haemolysis.

Further observations on nucleolar tails in amphibian oocytes

1991 ◽  
Vol 99 (3) ◽  
pp. 515-521
Author(s):  
PEDRO LEÓN ◽  
JAMES KEZER ◽  
ERIC SCHABTACH
Keyword(s):  
Electron Microscopy ◽  
Nuclear Envelope ◽  
Nuclear Membrane ◽  
Outer Surface ◽  
Light And Electron Microscopy ◽  
Nuclear Pores ◽  
Core Component ◽  
Amphibian Species ◽  
Attachment Structures ◽  
Convoluted Tubules

Large oocytes from some amphibian species possess beaded or unbeaded intranuclear tails that penetrate the extrachromosomal nucleoli through a distinct pit in their surface and attach to the central core component Here we show, using light and electron microscopy, that tails anchor nucleoli to the nuclear envelope through intricate attachment structures. These structures are composed of interconnected spherical masses containing highly convoluted tubules and associated extratubular proteins, directly directly in contact with the inner nuclear membrane. Fibers emerging from the nuclear pores seemingly hold the attachment complex in place. Beads on the nucleolar tails are formed by the accumulation of proteins on the outer surface of smooth tubules. The function of these intranuclear tubules is unknown

scholarly journals Exploring the Nuclear Envelope of Herpes Simplex Virus 1-Infected Cells by High-Resolution Microscopy

2008 ◽  
Vol 83 (1) ◽  
pp. 408-419 ◽  
Author(s):  
Peter Wild ◽  
Claudia Senn ◽  
Céline L. Manera ◽  
Esther Sutter ◽  
Elisabeth M. Schraner ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Herpes Simplex ◽  
Nuclear Envelope ◽  
Nuclear Pore ◽  
Nuclear Pores ◽  
Herpes Simplex Virus 1 ◽  
Infected Cells ◽  
The Mean ◽  
Simplex Virus ◽  
Hsv 1

ABSTRACT Herpesviruses are composed of capsid, tegument, and envelope. Capsids assemble in the nucleus and exit the nucleus by budding at the inner nuclear membrane, acquiring tegument and the envelope. This study focuses on the changes of the nuclear envelope during herpes simplex virus 1 (HSV-1) infection in HeLa and Vero cells by employing preparation techniques at ambient and low temperatures for high-resolution scanning and transmission electron microscopy and confocal laser scanning microscopy. Cryo-field emission scanning electron microscopy of freeze-fractured cells showed for the first time budding of capsids at the nuclear envelope at the third dimension with high activity at 10 h and low activity at 15 h of incubation. The mean number of pores was significantly lower, and the mean interpore distance and the mean interpore area were significantly larger than those for mock-infected cells 15 h after inoculation. Forty-five percent of nuclear pores in HSV-1-infected cells were dilated to more than 140 nm. Nuclear material containing capsids protrude through them into the cytoplasm. Examination of in situ preparations after dry fracturing revealed significant enlargements of the nuclear pore diameter and of the nuclear pore central channel in HSV-1-infected cells compared to mock-infected cells. The demonstration of nucleoporins by confocal microscopy also revealed fewer pores but focal enhancement of fluorescence signals in HSV-1-infected cells, whereas Western blots showed no loss of nucleoporins from cells. The data suggest that infection with HSV-1 alters the number, size, and architecture of nuclear pores without a loss of nucleoporins from altered nuclear pore complexes.

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).

New challenges to image processing posed by cryo-Electron microscopy of single particles

1991 ◽  
Vol 49 ◽  
pp. 422-423
Author(s):  
Joachim Frank
Keyword(s):  
Electron Microscopy ◽  
Phase Contrast ◽  
Particle Orientation ◽  
Single Particles ◽  
Contrast Transfer Function ◽  
Virtual Absence ◽  
Cryo Electron Microscopy ◽  
Spatial Frequencies ◽  
New Challenges ◽  
Particle Images

Compared with images of negatively stained single particle specimens, those obtained by cryo-electron microscopy have the following new features: (a) higher “signal” variability due to a higher variability of particle orientation; (b) reduced signal/noise ratio (S/N); (c) virtual absence of low-spatial-frequency information related to elastic scattering, due to the properties of the phase contrast transfer function (PCTF); and (d) reduced resolution due to the efforts of the microscopist to boost the PCTF at low spatial frequencies, in his attempt to obtain recognizable particle images.

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.

Fine structure and possible functions of the hermaphrodite duct of some pulmonate snails (Mollusca: Gastropoda)

1990 ◽  
Vol 48 (3) ◽  
pp. 68-69
Author(s):  
Alan N. Hodgson
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Fine Structure ◽  
Seminal Vesicle ◽  
Reproductive Biology ◽  
Light Microscope ◽  
Light Microscope Level ◽  
Transmission Electron ◽  
Standard Techniques

The hermaphrodite duct of pulmonate snails connects the ovotestis to the fertilization pouch. The duct is typically divided into three zones; aproximal duct which leaves the ovotestis, the middle duct (seminal vesicle) and the distal ovotestis duct. The seminal vesicle forms the major portion of the duct and is thought to store sperm prior to copulation. In addition the duct may also play a role in sperm maturation and degredation. Although the structure of the seminal vesicle has been described for a number of snails at the light microscope level there appear to be only two descriptions of the ultrastructure of this tissue. Clearly if the role of the hermaphrodite duct in the reproductive biology of pulmonatesis to be understood, knowledge of its fine structure is required.Hermaphrodite ducts, both containing and lacking sperm, of species of the terrestrial pulmonate genera Sphincterochila, Levantina, and Helix and the marine pulmonate genus Siphonaria were prepared for transmission electron microscopy by standard techniques.

High-voltage cytochemistry for three-dimensional observation of nuclei and calculation of total numbers of nuclear pores in retinal Mueller glia

1990 ◽  
Vol 48 (3) ◽  
pp. 956-957
Author(s):  
H. Ishigooka ◽  
S. Ueno ◽  
L.M. Hjelmeland ◽  
M.B. Landers ◽  
K. Ogawa
Keyword(s):  
Nuclear Envelope ◽  
High Voltage ◽  
Three Dimensional ◽  
Reaction Medium ◽  
Nuclear Pores ◽  
High Voltage Electron Microscopy ◽  
Voltage Electron ◽  
High Voltage Electron ◽  
G6pase Activity ◽  
Mueller Glia

Introduction: We have demonstrated that Glucose-6-phosphatase (G6Pase) activity is localized to the endoplasmic reticulum and nuclear envelope of Mueller glia in the normal and pathological guinea pig retina. Using a combination of this cytochemical technique and high voltage electron microscopy, the distribution of nuclear pores could be clearly observed on the nuclear envelope of Mueller glia because of their anatomical lack of reaction products. This technique was developed to study the three-dimensional structure of nuclei and to calculate total numbers of nuclear pores utilizing a computer graphic analysis system in the normal and pathological retina.Materials and methods: Normal and photocoagulated retina of pigmented adult guinea pigs were perfused with a cold mixture of 0.25% glutaraldehyde and 2% paraformaldehyde in 0.1M cacodylate buffer, and the enucleated globes were hemisected and immersed in the same fixative for 30 min. After sectioning and incubation in the reaction medium for the detection of G6Pase activity by the method of Wachstein-Meisel, the sections were postfixed, dehydrated and embedded in Spurr’s epoxy resin. Serial thick sections (1.0um) were prepared for the observation by a Hitachi high voltage electron microscope (H 1250-M) with an accelerating voltage of 1000 Kv. and pictures were analyzed and three-dimensionally reconstructed by TRI (RATOC Co., Ltd.).

Imaging sub-micron objects with light microscopy: Visualization of the T-4 bacteriophage

1989 ◽  
Vol 47 ◽  
pp. 834-835
Author(s):  
Malcolm Brown ◽  
Reynolds M. Delgado ◽  
Michael J. Fink
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Focal Plane ◽  
Phase Contrast ◽  
Dark Field ◽  
E Coli ◽  
Polystyrene Beads ◽  
Television Camera ◽  
Transmission Electron ◽  
Universal Microscope

While light microscopy has been used to image sub-micron objects, numerous problems with diffraction-limitations often preclude extraction of useful information. Using conventional dark-field and phase contrast light microscopy coupled with image processing, we have studied the following objects: (a) polystyrene beads (88nm, 264nm, and 557mn); (b) frustules of the diatom, Pleurosigma angulatum, and the T-4 bacteriophage attached to its host, E. coli or free in the medium. Equivalent images of the same areas of polystyrene beads and T-4 bacteriophages were produced using transmission electron microscopy.For light microscopy, we used a Zeiss universal microscope. For phase contrast observations a 100X Neofluar objective (N.A.=1.3) was applied. With dark-field, a 100X planachromat objective (N.A.=1.25) in combination with an ultra-condenser (N.A.=1.25) was employed. An intermediate magnifier (Optivar) was available to conveniently give magnification settings of 1.25, 1.6, and 2.0. The image was projected onto the back focal plane of a film or television camera with a Carl Zeiss Jena 18X Compens ocular.

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