scholarly journals THE NUCLEAR ANNULI AS PATHWAYS FOR NUCLEOCYTOPLASMIC EXCHANGES

1962 ◽  
Vol 14 (1) ◽  
pp. 65-72 ◽  
Author(s):  
Carl M. Feldherr
Keyword(s):  
Electron Microscopy ◽  
Nuclear Envelope ◽  
Colloidal Gold ◽  
Colloidal Particles ◽  
Time Interval ◽  
Central Channel ◽  
Time Intervals ◽  
Gold Particles ◽  
Pass Through ◽  
Individual Particles

Colloidal gold particles, 25 to 55 A in diameter, which had been coated with polyvinylpyrrolidone, were microinjected into the ground cytoplasm of amebas (Chaos chaos). At time intervals of 1 minute, 2 minutes, 10 minutes, and 24 hours after injection the cells were fixed for electron microscopy. After 24 hours, gold particles were found in both the nuclei and the ground cytoplasm, the concentration being higher in the nuclei. Colloidal particles were also present in the nuclei after 10 minutes, but at this time interval the concentration did not appear to be greater than that in the ground cytoplasm. One and 2 minutes after injection, the gold particles were located almost exclusively in the ground cytoplasm; however, individual particles were often found within the annuli of the nuclear envelope, and were located specifically in the centers of these structures. The results suggest that at least some of the gold particles which enter the nuclei pass through the annuli, and that passage through these structures may be restricted to a central channel.

Ultrastructural analysis of the spatial distribution of MRNA

1992 ◽  
Vol 50 (1) ◽  
pp. 552-553
Author(s):  
Gary Bassell ◽  
Robert H. Singer
Keyword(s):  
Electron Microscopy ◽  
Spatial Distribution ◽  
In Situ Hybridization ◽  
High Resolution ◽  
Nucleic Acids ◽  
Colloidal Gold ◽  
Dna Probe ◽  
Nucleotide Analogs ◽  
Gold Particles

We have been investigating the spatial distribution of nucleic acids intracellularly using in situ hybridization. The use of non-isotopic nucleotide analogs incorporated into the DNA probe allows the detection of the probe at its site of hybridization within the cell. This approach therefore is compatible with the high resolution available by electron microscopy. Biotinated or digoxigenated probe can be detected by antibodies conjugated to colloidal gold. Because mRNA serves as a template for the probe fragments, the colloidal gold particles are detected as arrays which allow it to be unequivocally distinguished from background.

Specificity of Lectins Labeled with Colloidal Gold to the Exopolymeric Matrix Carbohydrates of the Sulfate-Reducing Bacteria Biofilm Formed on Steel

2020 ◽  
Vol 82 (5) ◽  
pp. 11-20
Author(s):  
D.R. Abdulina ◽  
◽  
L.M. Purish ◽  
G.O. Iutynska ◽  
◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Colloidal Gold ◽  
Steel Surface ◽  
Lectin Binding ◽  
Sulfate Reducing Bacteria ◽  
Gold Particles ◽  
Sulfate Reducing ◽  
Transmission Electron ◽  
Reducing Bacteria

The studies of the carbohydrate composition of the sulfate-reducing bacteria (SRB) biofilms formed on the steel surface, which are a factor of microbial corrosion, are significant. Since exopolymers synthesized by bacteria could activate corrosive processes. The aim of the study was to investigate the specificity of commercial lectins, labeled with colloidal gold to carbohydrates in the biofilm exopolymeric matrix produced by the corrosive-relevant SRB strains from man-caused ecotopes. Methods. Microbiological methods (obtaining of the SRB biofilms during cultivation in liquid Postgate B media under microaerophilic conditions), biochemical methods (lectin-binding analysis of 10 commercial lectins, labeled with colloidal gold), transmission electron microscopy using JEM-1400 JEOL. Results. It was shown using transmission electron microscopy that the binding of lectins with carbohydrates in the biofilm of the studied SRB strains occurred directly in the exopolymerіс matrix, as well as on the surfaces of bacterial cells, as seen by the presence of colloidal gold particles. For detection of the neutral carbohydrates (D-glucose and D-mannose) in the biofilm of almost all studied bacterial strains PSA lectin was the most specific. This lectin binding in biofilms of Desulfotomaculum sp. К1/3 and Desulfovibrio sp. 10 strains was higher in 90.8% and 94.4%, respectively, then for ConA lectin. The presence of fucose in the SRB biofilms was detected using LABA lectin, that showed specificity to the biofilm EPS of all the studied strains. LBA lectin was the most specific to N-аcetyl-D-galactosamine for determination of amino sugars in the biofilm. The amount of this lectin binding in D. vulgaris DSM644 biofilm was 30.3, 10.1 and 9.3 times higher than SBA, SNA and PNA lectins, respectively. STA, LVA and WGA lectins were used to detect the N-acetyl-Dglucosamine and sialic acid in the biofilm. WGA lectin showed specificity to N-acetyl-D-glucosamine in the biofilm of all the studied SRB; maximum number of bounded colloidal gold particles (175 particles/μm2) was found in the Desulfotomaculum sp. TC3 biofilm. STA lectin was interacted most actively with N-acetyl-D-glucosamine in Desulfotomaculum sp. TC3 and Desulfomicrobium sp. TC4 biofilms. The number of bounded colloidal gold particles was in 9.2 and 7.4 times higher, respectively, than using LVA lectin. The lowest binding of colloidal gold particles was observed for LVA lectin. Conclusions. It was identified the individual specificity of the 10 commercial lectins to the carbohydrates of biofilm matrix on the steel surface, produced by SRB. It was estimated that lectins with identical carbohydrates specificity had variation in binding to the biofilm carbohydrates of different SRB strains. Establishing of the lectin range selected for each culture lead to the reduction of the scope of studies and labor time in the researching of the peculiarities of exopolymeric matrix composition of biofilms formed by corrosiverelevant SRB.

Colloidal gold particles as a new in vivo marker of early acute lung injury

2004 ◽  
Vol 287 (4) ◽  
pp. L867-L878 ◽  
Author(s):  
Kai Heckel ◽  
Rainer Kiefmann ◽  
Martina Dörger ◽  
Mechthild Stoeckelhuber ◽  
Alwin E. Goetz
Keyword(s):  
Electron Microscopy ◽  
Acute Lung Injury ◽  
Gas Exchange ◽  
Lung Injury ◽  
Colloidal Gold ◽  
Microvascular Permeability ◽  
Arterial Oxygen ◽  
Control Group ◽  
Gold Particles

Permeability of the endothelial barrier to large molecules plays a pivotal role in the manifestation of early acute lung injury. We present a novel and sensitive technique that brings microanatomical visualization and quantification of microvascular permeability in line. White New Zealand rabbits were anesthetized and ventilated mechanically. Rabbit serum albumin (RSA) was labeled with colloidal gold particles. We quantified macromolecular leakage of gold-labeled RSA and thickening of the gas exchange distance by electron microscopy, taking into account morphology of microvessels. The control group receiving a saline solution represented a normal gas exchange barrier without extravasation of gold-labeled albumin. Infusion of lipopolysaccharide (LPS) resulted in a significant displacement of gold-labeled albumin into pulmonary cells, the lung interstitium, and even the alveolar space. Correspondingly, intravital fluorescence microscopy and digital image analysis indicated thickening of width of alveolar septa. The findings were accompanied by a deterioration of alveolo-arterial oxygen difference, whereas wet/dry ratio and albumin concentration in the bronchoalveolar lavage fluid failed to detect that early stage of pulmonary edema. Inhibition of the nuclear enzyme poly(ADP-ribose) synthetase by 3-aminobenzamide prevented LPS-induced microvascular injury. To summarize: colloidal gold particles visualized by standard electron microscopy are a new and very sensitive in vivo marker of microvascular permeability in early acute lung injury. This technique enabling detailed microanatomical and quantitative pathophysiological characterization of edema formation can form the basis for evaluating novel treatment strategies against acute lung injury.

scholarly journals Preparation of Protein a Gold

1997 ◽  
Vol 5 (5) ◽  
pp. 12-13
Author(s):  
Paul Webster
Keyword(s):  
Electron Microscopy ◽  
Biological Activity ◽  
Colloidal Gold ◽  
Protein A ◽  
List Type ◽  
Type Number ◽  
Stained Glass ◽  
Small Particles ◽  
Silver Enhancement ◽  
Gold Particles

Colloidal gold has been used for centuries in the preparation of stained glass for windows and fine glassware. In recent years, colloidal gold particles have become a useful tool in microscopy for staining tissues and sections. Colloidal gold particles are especially useful for biological electron microscopy, Some of the reasons why are listed below.*Homogeneous preparations of particles varying in size from 3μm to 20μm can be easily prepared.*Colloidal gold suspensions are inexpensive to prepare. Most proteins can be easily coupled to colloidal gold particles.*Most proteins can be easily coupled to colloidal gold particles.*Proteins coupled to gold particles do not appear to lose their biological activity.*The colloidal gold particles can be easily seen in the electron microscope.*Colloidal gold does not naturally occur in biological material. Therefore, if you see it, it is because you put it there.*Colloidal gold probes can be used for light microscopy, The larger gold particles can be directly observed by the light microscope. Small particles are detected by silver enhancement or epipolarized illumination.*The same probes can be used for both LM and TEM imrnunocytochemistry.

scholarly journals THE EFFECT OF THE ELECTRON-OPAQUE PORE MATERIAL ON EXCHANGES THROUGH THE NUCLEAR ANNULI

1965 ◽  
Vol 25 (1) ◽  
pp. 43-53 ◽  
Author(s):  
Carl M. Feldherr
Keyword(s):  
Particle Size ◽  
Electron Microscope ◽  
Nuclear Envelope ◽  
Colloidal Gold ◽  
Maximum Size ◽  
Amoeba Proteus ◽  
Gold Particles ◽  
High Concentrations

To investigate the extent to which the electron-opaque pore material can regulate nucleocytoplasmic exchanges which occur through the nuclear annuli, experiments were performed in which polyvinylpyrrolidone (PVP)-coated colloidal gold particles (25 to 170 A in diameter) were microinjected into the cytoplasm of amebas (Amoeba proteus). The cells were fixed at various times after injection and examined with the electron microscope in order to determine the location of the gold particles. High concentrations of gold were found associated with the pore material at specific points adjacent to and within the pores. It is tentatively suggested that such specific accumulations could be a means of selecting substances from the cytoplasm for transport through the pores. Particles were also scattered throughout the ground cytoplasm and nucleoplasm. A comparison of the diameters of particles located in these two regions showed that the ability of materials to penetrate the nuclear envelope is a function of their size. It was estimated that the maximum size of the particles able to enter the nucleus is approximately 125 to 145 A indiameter. The regulation of exchanges with regard to particle size is thought to be dependent on the specific organization of the electron-opaque pore material.

scholarly journals Ultracytochemistry of calmodulin binding sites in myocardial cells by staining of frozen thin sections with colloidal gold-labeled calmodulin.

1989 ◽  
Vol 37 (2) ◽  
pp. 249-256 ◽  
Author(s):  
K Fujimoto ◽  
N Araki ◽  
K S Ogawa ◽  
S Kondo ◽  
T Kitaoka ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Sarcoplasmic Reticulum ◽  
Binding Sites ◽  
Colloidal Gold ◽  
Biologically Active ◽  
Myocardial Cells ◽  
Thin Sections ◽  
Gold Particles ◽  
Calmodulin Binding ◽  
Tissue Sections

Calmodulin (CaM) has been implicated as a multifunctional regulator of Ca2+ in the cytoplasm of cells. We have recently introduced biologically active colloidal gold-labeled CaM as a marker for identifying potential CaM binding sites (unoccupied by endogenous CaM at the time of fixation) by electron microscopy and have stained frozen thin sections of rat cardiac muscle with this conjugate. In the presence of Ca2+, gold particles indicating CaM binding sites were found localized on the sarcoplasmic reticulum, mitochondria, and gap junctions. Control tissue sections treated with EGTA or exposed to excess amounts of unlabeled native CaM before staining showed no binding. We believe that cytochemistry of potential CaM binding sites revealed by staining with labeled exogenous CaM is useful in correlating known biochemical reactions of CaM with particular cell activities.

Colloidal gold double labeling of two secretory proteins in bacterial cell

1983 ◽  
Vol 41 ◽  
pp. 614-615
Author(s):  
Guan Tinglu ◽  
Arati Ghosh ◽  
Bijan K. Ghosh
Keyword(s):  
Alkaline Phosphatase ◽  
Colloidal Gold ◽  
Colloidal Particles ◽  
Sodium Citrate ◽  
Secretory Proteins ◽  
Bacterial Cells ◽  
Double Labeling ◽  
Antibody Technique ◽  
Gold Particles ◽  
Multiple Labelling

Secretory proteins have been localized in bacterial cells by inmunoelectron-microscopy and cytochemical methods. Although ferritin-antibody technique is satisfactory for the localization of single antigenic sites, multiple labelling is not possible by this technique. However, such multiple labelling is possible using colloidal gold antibody because colloidal gold of widely varying sizes can be prepared by simple laboratory manipulation. We prepared colloidal gold of two different sizes and labelled these with anti-penicillinase and anti-alkaline phosphatase antibodies. These labels were used to determine the location of penicillinase and alkaline phosphatase in secreting Bacillus licheniformis 749/C cells.Large colloidal gold particles (150Å) were prepared by 10% sodium citrate treatment of 0.01% HAuCl4; the pH of this colloidal gold suspension was adjusted to 7.6 with 0.2 M K2CO3. Small colloidal particles (50A) were prepared by treating 0.01% HAuCl4, after pH was adjusted to 8.0 with 0.2M K2CO3, withphosphorous saturated ether. The large or small gold particles (1010 in 10 ml) were treated with 100μg of anti-alkaline phosphatase or anti-penicillinase antibodies; after thorough mixing unbound antibodies were removed by centrifugation.

scholarly journals ELECTRON MICROSCOPY OF HELA CELLS AFTER THE INGESTION OF COLLOIDAL GOLD

1957 ◽  
Vol 3 (5) ◽  
pp. 749-756 ◽  
Author(s):  
Carl G. Harford ◽  
Alice Hamlin ◽  
Esther Parker
Keyword(s):  
Electron Microscopy ◽  
Endoplasmic Reticulum ◽  
Cell Membrane ◽  
Golgi Complex ◽  
Hela Cells ◽  
Colloidal Gold ◽  
Tissue Cultures ◽  
Cytoplasmic Matrix ◽  
Gold Particles ◽  
Dense Granules

Tissue cultures of HeLa cells were grown in media containing colloidal gold, and after various intervals, the cells were fixed, embedded, and sectioned for electron microscopy. Uncoated grids with small holes were used in many of the experiments. Intracellular particles of gold were identified in areas surrounded by single membranes, in moderately dense granules, in globoid bodies, and in the cytoplasmic matrix. Gold particles were not found in typical mitochondria, Golgi complex, ergastoplasm (granular forms of endoplasmic reticulum), or nuclei. The phenomenon of pinocytosis was considered to be the most likely means by which the gold particles were ingested, and the locations of gold particles appeared to have significance concerning theories that membranous organelles of the cytoplasm may be derived from the cell membrane.

scholarly journals Localization of immunoreactive tyrosine hydroxylase in the goldfish retina with pre-embedding immunolabeling with one-nanometer colloidal gold particles and gold toning.

1992 ◽  
Vol 40 (10) ◽  
pp. 1465-1470 ◽  
Author(s):  
D W Marshak
Keyword(s):  
Electron Microscopy ◽  
Tyrosine Hydroxylase ◽  
Colloidal Gold ◽  
Light And Electron Microscopy ◽  
Rabbit Antiserum ◽  
Gold Chloride ◽  
Thin Sections ◽  
Gold Particles ◽  
Rabbit Igg ◽  
Goldfish Retina

The goal of this study was to develop an alternative to silver intensification for visualizing small colloidal gold particles by light and electron microscopy. The isolated goldfish retina was labeled with rabbit antiserum to tyrosine hydroxylase and 1-nm colloidal gold-conjugated goat anti-rabbit IgG. The gold particles were enlarged by toning with gold chloride, followed by reduction in oxalic acid. Dopaminergic interplexiform cells were clearly visible by light microscopy and, in lightly-fixed material treated with detergent, they were labeled in their entirety. Labeling was qualitatively similar, although less extensive, in material fixed and processed for electron microscopy. The labeled processes were apparent in ultra-thin sections viewed at low magnification, but the gold-toned particles were not so large that they obscured subcellular structures. The procedure apparently had no deleterious effects on the tissue, since the ultrastructural preservation was comparable to that seen with other pre-embedding immunolabeling methods. The technique was simple, reliable and, since the gold solutions were so dilute, relatively inexpensive.

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