Single Processing of Tissue for Light and Electron Microscopy

1977 ◽  
Vol 35 ◽  
pp. 550-551 ◽  
Author(s):  
W. A. Burns ◽  
A. M. Bretschneider ◽  
A. B. Morrison
Keyword(s):  
Electron Microscopy ◽  
Light And Electron Microscopy ◽  
Ultrastructural Level ◽  
Significant Problem ◽  
Diagnostic Ability ◽  
Large Samples ◽  
Microscopic Evaluation ◽  
Light Microscopist ◽  
Plastic Embedding ◽  
Electron Microscopist

Tissues for light and electron microscopy are traditionally processed separately. Different fixatives and embedding media are usually employed. Paraffin is unsuitable for electron microscopy and the small amounts of tissue generally embedded in plastic makes sampling a significant problem for the light, as well as the electron microscopist. Techniques, however, have been described for plastic embedding of large samples of tissue which can be sectioned at 1 μm. The added resolution of these thinner sections potentially increases the diagnostic ability of the light microscopist. These techniques have not been fully utilized in pathology. If one fixative were utilized and the quantity embedded in plastic were comparable to that which is normally processed for paraffin, then the investigator could use plastic sections for better light microscopic evaluation with the option of subsequent examination of the same region in the same block at an ultrastructural level.

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.

scholarly journals A microwave method for plastic embedding of nervous tissue for light and electron microscopy

Heliyon ◽  
2020 ◽  
Vol 6 (1) ◽  
pp. e03036
Author(s):  
Evan Calkins ◽  
Edvinas Pocius ◽  
Gail Marracci ◽  
Priya Chaudhary
Keyword(s):  
Electron Microscopy ◽  
Nervous Tissue ◽  
Light And Electron Microscopy ◽  
Microwave Method ◽  
Plastic Embedding

Efficient fluorescence recovery using antifade reagents in correlative light and electron microscopy

Microscopy ◽  
2019 ◽  
Vol 68 (5) ◽  
pp. 417-421
Author(s):  
Kiminori Toyooka ◽  
Naeko Shinozaki-Narikawa
Keyword(s):  
Electron Microscopy ◽  
Fluorescent Protein ◽  
Light And Electron Microscopy ◽  
Ultrastructural Level ◽  
Fluorescence Decay ◽  
Fluorescence Recovery ◽  
Recovery Method ◽  
Thin Sections ◽  
Electron Microscopies ◽  
Green Fluorescent

Abstract Correlative light and electron microscopy (CLEM) enables ultrastructural-level analysis of fluorescence-labeled proteins by combining images obtained from both fluorescence and electron microscopies. A technical challenge with the CLEM method is the effective detection of fluorescence from samples embedded in resins, which generally cause fluorescence decay. To overcome this issue, we developed a method for fluorescence recovery of green fluorescent protein (GFP) in resin-embedded semi-thin sections using commercially available antifade reagents. By applying this method, we successfully obtained CLEM images using field-emission scanning electron microscopy with moderately enhanced GFP signals, demonstrating the efficacy of this simple fluorescence recovery method.

scholarly journals Ultrastructural Dendritic Changes Underlying Diaschisis After Capsular Infarct

2020 ◽  
Vol 79 (5) ◽  
pp. 508-517
Author(s):  
Min-Cheol Lee ◽  
Ra Gyung Kim ◽  
Taebum Lee ◽  
Jo-Heon Kim ◽  
Kyung-Hwa Lee ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Light And Electron Microscopy ◽  
Ultrastructural Level ◽  
Subcortical White Matter ◽  
Ultrastructural Changes ◽  
Pathological Changes ◽  
Hydropic Degeneration ◽  
Mild Degree ◽  
Neuronal Soma ◽  
Axodendritic Synapses

Abstract Diaschisis has been described as functional depression distant to the lesion. A variety of neuroscientific approaches have been used to investigate the mechanisms underlying diaschisis. However, few studies have examined the pathological changes in diaschisis at ultrastructural level. Here, we used a rat model of capsular infarct that consistently produces diaschisis in ipsilesional and contralesional motor and sensory cortices. To verify the occurrence of diaschisis and monitor time-dependent changes in diaschisis, we performed longitudinal 2-deoxy-2-[18F]-fluoro-d-glucose microPET (FDG-microPET) study. We also used light and electron microscopy to identify the microscopic and ultrastructural changes at the diaschisis site at 7, 14, and 21 days after capsular infarct modeling (CIM). FDG-microPET showed the occurrence of diaschisis after CIM. Light microscopic examinations revealed no significant histopathological changes at the diaschisis site except a mild degree of reactive astrogliosis. However, electron microscopy revealed swollen, hydropic degeneration of axial dendrites and axodendritic synapses, although the neuronal soma (including nuclear chromatin and cytoplasmic organelles) and myelinated axons were relatively well preserved up to 21 days after injury. Furthermore, number of axodendritic synapses was significantly decreased after CIM. These data indicate that a circumscribed subcortical white-matter lesion produces ultrastructural pathological changes related to the pathogenesis of diaschisis.

scholarly journals CLEMSite, a software for automated phenotypic screens using light microscopy and FIB-SEM

2021 ◽  
Author(s):  
José M. Serra Lleti ◽  
Anna M. Steyer ◽  
Nicole L. Schieber ◽  
Beate Neumann ◽  
Christian Tischer ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Light Microscopy ◽  
Focused Ion Beam ◽  
Cultured Cells ◽  
Ion Beam ◽  
Light And Electron Microscopy ◽  
Ultrastructural Level ◽  
Reference Points ◽  
Grid Pattern ◽  
Volume Acquisition

AbstractCorrelative light and electron microscopy (CLEM) combines two imaging modalities, balancing out the limits of one technique with the other. In recent years, Focused Ion Beam Scanning Electron Microscopy (FIB-SEM) has emerged as a flexible method that enables semi-automated volume acquisition at the ultrastructural level. We present a toolset for adherent cultured cells that enables tracking and finding cell regions previously identified in light microscopy, in the FIB-SEM along with automatic acquisition of high-resolution volume datasets. We detect a grid pattern in both modalities (LM and EM), which identifies common reference points. The novel combination of these techniques enables complete automation of the workflow. This includes setting the coincidence point of both ion and electron beams, automated evaluation of the image quality and constantly tracking the sample position with the microscope’s field of view reducing or even eliminating operator supervision. We show the ability to target the regions of interest in EM within 5µm accuracy, while iterating between different targets including unattended data acquisition. Our results demonstrate that executing high throughput volume acquisition in electron microscopy is possible.

Feulgen-thiocarbohydrazide-silver methenamine(FTS) reaction: An improved silver methodology for light and electron microscopy of DNA

1984 ◽  
Vol 42 ◽  
pp. 262-263 ◽  
Author(s):  
J. S. Hanker ◽  
B. L. Giammara
Keyword(s):  
Electron Microscopy ◽  
High Resolution ◽  
Molecular Species ◽  
Light And Electron Microscopy ◽  
Ultrastructural Level ◽  
Feulgen Reaction ◽  
Full Paper ◽  
End Products ◽  
Silver Compounds

A number of cytochemical methods have been Introduced to reveal nuclear components at the ultrastructural level. Many of these, however, are topographic, i.e., identify structures rather than molecular species (1). Indeed, even the search for modifications of the Feulgen reaction to render the nuclear sites of chromatin or DNA electron opaque have not met with a great deal of success (1). Thus the aldehyde or pseudoaldehyde groups on apurinic acids, generated by the action of IN HCl at 60° on DNA, could not be shown by condensation with thiocarbohydrazide (TCH) or thiosemicarbazide and subsequent osmication (2,3). Feulgen reactions with silver end products resulting from the reaction of silver compounds with apurinic-acids have been used since 1924, but these reactions either lack specificity or cannot be used for high resolution studies. Although an abstract appeared in 1972 on a Feulgen reaction employing TCH and silver proteinate (4), neither light micrographs nor a full paper have appeared.

Cytological study of Сhloromonas typhlos (Chlamydomonadaceae, Chlorophyta) from North-Western Russia

2017 ◽  
Vol 51 ◽  
pp. 5-11 ◽  
Author(s):  
O. N. Boldina
Keyword(s):  
Electron Microscopy ◽  
Cytological Study ◽  
Light And Electron Microscopy ◽  
First Record ◽  
Ultrastructural Level ◽  
North Western

The strain of biflagellated green monad, selected from the specimen gathered in the Novgorod Region was studied by light and electron microscopy. The species was identified on the base of coincidence of the majority of specific light and ultrastructural signs revealed in both the strain SAG 26.86 of Chloromonas typhlos (Gerloff) Matsuzaki et al. and in studied strain. On LM level the cells are ellipsoid, some asymmetric, 12–17 μm long, 8–13 μm wide, with an apical, hardly distinguished hemispherical papilla flattened on the top. Chloroplast parietal, cup-shaped, thickened on one side, with big lobes, irregular incisions and fissures on the surface. These and other features are characteristic of the type and the studied strain of C. typhlos, excluding stigma observed in the studied strain only. On the ultrastructural level, large thylakoid packs, similar with chloroplast thylakoids, were observed in pyrenoids of both strains of C. typhlos. The detected pyrenoid type was never discovered in chlamydomonads earlier. It is the first record of C. typhlos for the Novgorod Region and the whole Russia, and adds the list of «bloom-forming» species. The strain was deposited in the collection of microalgae strains in the Institute of Biology of Komi Scientific Centre under the name SYKOA Ch-063-17.

A rapid silver stain for the DNA of microorganisms cultured from AIDS and other immunocompromised patients

1990 ◽  
Vol 48 (3) ◽  
pp. 762-763 ◽  
Author(s):  
B.L Giammara ◽  
R.L. Hopfer ◽  
P.E. Yates ◽  
J.S. Hanker
Keyword(s):  
Electron Microscopy ◽  
Light And Electron Microscopy ◽  
Subsequent Development ◽  
Ultrastructural Level ◽  
Nuclear Chromatin ◽  
Immunocompromised Patients ◽  
Feulgen Reaction ◽  
Silver Stain ◽  
Mw Irradiation ◽  
Current Procedure

Although a number of cytochemical methods have been introduced for nuclei at the ultrastructural level, many of these are topographic, i.e. identify nuclei rather than nuclear chromatin and DNA. An improved silver method for the Feulgen reaction for the light and electron microscopy of DNA was introduced in 1984 (FETS reaction). It required application of the silver methenamine for 15 min. under UV-irradiation and subsequent development of the stain overnight in glycerine. This type of methodology has been considerably accelerated by the application of silver solutions, especially silver methenamine solutions prepared from the solid, under microwave(MW) irradiation. The current procedure employs 1 min. formalin fixation of a smear of the sample on a slide or coverslip. The fixed smear is then rinsed 1 hr. in buffer. Aldehyde or pseudoaldehyde groups of apurinic acids are generated by the action of 2.5N HCl for 10 min. on the DNA. These groups are then condensed with thiocarbohydrazide and the resulting product reacted with silver methenamine solution under MW irradiation for 4 min.

scholarly journals Correlative Light and Electron Microscopy (CLEM) Analysis of Nuclear Reorganization Induced by Clustered DNA Damage Upon Charged Particle Irradiation

2020 ◽  
Vol 21 (6) ◽  
pp. 1911 ◽  
Author(s):  
Susanne Tonnemacher ◽  
Mikhail Eltsov ◽  
Burkhard Jakob
Keyword(s):  
Electron Microscopy ◽  
Dna Damage ◽  
Light And Electron Microscopy ◽  
Ultrastructural Level ◽  
Uranyl Acetate ◽  
Dna Double Strand Breaks ◽  
Nuclear Reorganization ◽  
Particle Irradiation ◽  
Transmission Electron ◽  
Radiation Induced

Chromatin architecture plays major roles in gene regulation as well as in the repair of DNA damaged by endogenous or exogenous factors, such as after radiation. Opening up the chromatin might provide the necessary accessibility for the recruitment and binding of repair factors, thus facilitating timely and correct repair. The observed formation of ionizing radiation-induced foci (IRIF) of factors, such as 53BP1, upon induction of DNA double-strand breaks have been recently linked to local chromatin decompaction. Using correlative light and electron microscopy (CLEM) in combination with DNA-specific contrasting for transmission electron microscopy or tomography, we are able to show that at the ultrastructural level, these DNA damage domains reveal a chromatin compaction and organization not distinguishable from regular euchromatin upon irradiation with carbon or iron ions. Low Density Areas (LDAs) at sites of particle-induced DNA damage, as observed after unspecific uranyl acetate (UA)-staining, are thus unlikely to represent pure chromatin decompaction. RNA-specific terbium-citrate (Tb) staining suggests rather a reduced RNA density contributing to the LDA phenotype. Our observations are discussed in the view of liquid-like phase separation as one of the mechanisms of regulating DNA repair.

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