Further characterization of the type 3 ryanodine receptor (RyR3) purified from rabbit diaphragm. (1/80)

We characterized type 3 ryanodine receptor (RyR3) purified from rabbit diaphragm by immunoaffinity chromatography using a specific antibody. The purified receptor was free from 12-kDa FK506-binding protein, although it retained the ability to bind 12-kDa FK506-binding protein. Negatively stained images of RyR3 show a characteristic rectangular structure that was indistinguishable from RyR1. The location of the D2 segment, which exists uniquely in the RyR1 isoform, was determined as the region around domain 9 close to the corner of the square-shaped assembly, with use of D2-directed antibody as a probe. The RyR3 homotetramer had a single class of high affinity [3H]ryanodine-binding sites with a stoichiometry of 1 mol/mol. In planar lipid bilayers, RyR3 displayed cation channel activity that was modulated by several ligands including Ca2+, Mg2+, caffeine, and ATP, which is consistent with [3H]ryanodine binding activity. RyR3 showed a slightly larger unit conductance and a longer mean open time than RyR1. Whereas RyR1 showed two classes of channel activity with distinct open probabilities (Po), RyR3 displayed a homogeneous and steeply Ca2+-dependent activity with Po approximately 1. RyR3 was more steeply affected in the channel activity by sulfhydryl-oxidizing and -reducing reagents than RyR1, suggesting that the channel activity of RyR3 may be transformed more precipitously by the redox state. This is also a likely explanation for the difference in the Ca2+ dependence of RyR3 between [3H]ryanodine binding and channel activity.  (+info)

Dose-dependent linkage, assembly inhibition and disassembly of vimentin and cytokeratin 5/14 filaments through plectin's intermediate filament-binding domain. (2/80)

Plectin, the largest and most versatile member of the cytolinker/plakin family of proteins characterized to date, has a tripartite structure comprising a central 200 nm-long (&agr;)-helical rod domain flanked by large globular domains. The C-terminal domain comprises a short tail region preceded by six highly conserved repeats (each 28-39 kDa), one of which (repeat 5) contains plectin's intermediate filament (IF)-binding site. We used recombinant and native proteins to assess the effects of plectin repeat 5-binding to IF proteins of different types. Quantitative Eu(3+)-based overlay assays showed that plectin's repeat 5 domain bound to type III IF proteins (vimentin) with preference over type I and II cytokeratins 5 and 14. The ability of both types of IF proteins to self-assemble into filaments in vitro was impaired by plectin's repeat 5 domain in a concentration-dependent manner, as revealed by negative staining and rotary shadowing electron microscopy. This effect was much more pronounced in the case of vimentin compared to cytokeratins 5/14. Preassembled filaments of both types became more and more crosslinked upon incubation with increasing concentrations of plectin repeat 5. However, at high proportions of plectin to IF proteins, disassembly of filaments occurred. Again, vimentin filaments proved considerably more sensitive towards disassembly than those composed of cytokeratins 5 and 14. In general, IFs formed from recombinant proteins were found to be slightly more responsive towards plectin influences than their native counterparts. A dose-dependent plectin-inflicted collapse and putative disruption of IFs was also observed in vivo after ectopic expression of vimentin and plectin's repeat 5 domain in cotransfected vimentin-deficient SW13 (vim(-)) cells. Our results suggest an involvement of plectin not only in crosslinking and stabilization of cytoskeletal IF networks, but also in regulation of their dynamics.  (+info)

Nonsaturable binding indicates clustering of tau on the microtubule surface in a paired helical filament-like conformation. (3/80)

Tau protein modulates microtubule dynamics and forms insoluble aggregates in Alzheimer's disease. Because there is a discrepancy between reported affinities of Tau to microtubules, we determined the interaction over a wide concentration range using a sensitive enzyme-linked immunosorbent assay. We found that the interaction is biphasic and not monophasic as assumed earlier. The first binding phase is typical for identical and noninteracting binding sites, with dissociation constants around 0.1 micrometer and stoichiometries around 0.2 Tau/tubulin dimer. Surprisingly, the second phase is nonsaturable and shows a nearly linear increase in bound Tau versus free Tau for free Tau concentrations higher than 2 micrometer. The slope is proportional to the microtubule concentration. From this we define an overloading parameter with values around 50 micrometer. The influence of Tau isoform, phosphorylation, and dimerization on both phases was investigated. Remarkably the overloading of Tau on microtubules leads to a thioflavin S fluorescence increase reminiscent of that seen with Tau aggregated into Alzheimer paired helical filaments. Because polyanions stimulate Tau aggregation and because the C-terminal domain of tubulin is polyanionic, we suggest that an early conformational change in Tau leading to paired helical filament aggregation occurs right on the microtubule surface.  (+info)

Novel filaments 5 nm in diameter constitute the cytosolic ring of the plastid division apparatus. (4/80)

The plastid division apparatus (called the plastid-dividing ring) has been detected in several plant and algal species at the constricted region of plastids by transmission electron microscopy. The apparatus is composed of two or three rings: an outer ring in the cytosol, an inner ring in the stroma, and a middle ring in the intermembrane space. The components of these rings are not clear. FtsZ, which forms the bacterial cytokinetic ring, has been proposed as a component of both the inner and outer rings. Here, we present the ultrastructure of the outer ring at high resolution. To visualize the outer ring by negative staining, we isolated dividing chloroplasts from a synchronized culture of a red alga, Cyanidioschyzon merolae, and lysed them with nonionic detergent Nonidet P-40. Nonidet P-40 extracted primarily stroma, thylakoids, and the inner and middle rings, leaving the envelope and outer ring largely intact. Negative staining revealed that the outer ring consists of a bundle of 5-nm filaments in which globular proteins are spaced 4.8 nm apart. Immunoblotting using an FtsZ-specific antibody failed to show immunoreactivity in the fraction containing the filament. Moreover, the filament structure and properties are unlike those of known cytoskeletal filaments. The bundle of filaments forms a very rigid structure and does not disassemble in 2 M urea. We also identified a dividing phase-specific 56-kD protein of chloroplasts as a candidate component of the ring. Our results suggest that the main architecture of the outer ring did not descend from cyanobacteria during the course of endosymbiosis but was added by the host cell early in plant evolution.  (+info)

Streptococcus salivarius fimbriae are composed of a glycoprotein containing a repeated motif assembled into a filamentous nondissociable structure. (5/80)

Streptococcus salivarius, a gram-positive bacterium found in the human oral cavity, expresses flexible peritrichous fimbriae. In this paper, we report purification and partial characterization of S. salivarius fimbriae. Fimbriae were extracted by shearing the cell surface of hyperfimbriated mutant A37 (a spontaneous mutant of S. salivarius ATCC 25975) with glass beads. Preliminary experiments showed that S. salivarius fimbriae did not dissociate when they were incubated at 100 degrees C in the presence of sodium dodecyl sulfate. This characteristic was used to separate them from other cell surface components by successive gel filtration chromatography procedures. Fimbriae with molecular masses ranging from 20 x 10(6) to 40 x 10(6) Da were purified. Examination of purified fimbriae by electron microscopy revealed the presence of filamentous structures up to 1 microm long and 3 to 4 nm in diameter. Biochemical studies of purified fimbriae and an amino acid sequence analysis of a fimbrial internal peptide revealed that S. salivarius fimbriae were composed of a glycoprotein assembled into a filamentous structure resistant to dissociation. The internal amino acid sequence was composed of a repeated motif of two amino acids alternating with two modified residues: A/X/T-E-Q-M/phi, where X represents a modified amino acid residue and phi represents a blank cycle. Immunolocalization experiments also revealed that the fimbriae were associated with a wheat germ agglutinin-reactive carbohydrate. Immunolabeling experiments with antifimbria polyclonal antibodies showed that antigenically related fimbria-like structures were expressed in two other human oral streptococcal species, Streptococcus mitis and Streptococcus constellatus.  (+info)

Analysis of the PilQ secretin from Neisseria meningitidis by transmission electron microscopy reveals a dodecameric quaternary structure. (6/80)

PilQ is a member of the secretin family of outer membrane proteins and is specifically involved in secretion of type IV pili in Neisseria meningitidis, Neisseria gonorrhoeae, and Pseudomonas aeruginosa. The quaternary structure of PilQ from N. meningitidis was analyzed by transmission electron microscopy by using a negative stain. Single particle averaging was carried out with a total data set of 650 individual particles, which produced a projection map generated from 296 particles at an estimated resolution of 2.6 nm. Oligomeric PilQ adopts a donut-like structure with an external ring that is 16.5 nm in diameter surrounding a central cavity that is 6.5 nm in diameter. Self-rotation and power spectrum analysis demonstrated the presence of 12-fold rotational symmetry, showing that PilQ is organized as a ring of 12 identical subunits. A model of the type IV meningococcal pilus fiber, based on the X-ray crystal structure of the N. gonorrhoeae pilin subunit, fitted neatly into the cavity, demonstrating how PilQ could serve as a channel for the growing pilus fiber.  (+info)

Three-dimensional architecture of the eukaryotic multisynthetase complex determined from negatively stained and cryoelectron micrographs. (7/80)

This study provides the first description of the three-dimensional architecture of the multienzyme complex of aminoacyl-tRNA synthetases. Reconstructions were calculated from electron microscopic images of negatively stained and frozen hydrated samples using three independent angular assignment methods. In all cases, volumes show an asymmetric triangular arrangement of protein domains around a deep central cavity. The structures have openings or indentations on most sides. Maximum dimensions are ca. 19x16x10 nm. The central cavity is 4 nm in diameter and extends two-thirds of the length of the particle.  (+info)

Projection structure of the photosynthetic reaction centre-antenna complex of Rhodospirillum rubrum at 8.5 A resolution. (8/80)

Two-dimensional crystals of the reaction-centre-light-harvesting complex I (RC-LH1) of the purple non- sulfur bacterium Rhodospirillum rubrum have been formed from detergent-solubilized and purified protein complexes. Unstained samples of this intrinsic membrane protein complex have been analysed by electron cryomicroscopy (cryo EM). Projection maps were calculated to 8.5 A from two different crystal forms, and show a single reaction centre surrounded by 16 LH1 subunits in a ring of approximately 115 A diameter. Within each LH1 subunit, densities for the alpha- and beta-polypeptide chains are clearly resolved. In one crystal form the LH1 forms a circular ring, and in the other form the ring is significantly ellipsoidal. In each case, the reaction centre adopts preferred orientations, suggesting specific interactions between the reaction centre and LH1 subunits rather than a continuum of possible orientations with the antenna ring. This experimentally determined structure shows no evidence of any other protein components in the closed LH1 ring. The demonstration of circular or elliptical forms of LH1 indicates that this complex is likely to be flexible in the bacterial membrane.  (+info)

"Negative staining for dummies". Retrieved 2009-06-06. "Negative staining". Archived from the original on 2012-12-25. Retrieved ... This contrasts with positive staining, in which the actual specimen is stained. For bright-field microscopy, negative staining ... The choice of negative stain in electron microscopy can be very important. An early study of plant viruses using negatively ... Negative staining is usually a very mild preparation method and thus does not reduce the possibility of operator infection. ...
Because only one stain is being used, the specimens (for positive stains) or background (for negative stains) will be one color ... Common variants include Wright's stain, Jenner's stain, May-Grunwald stain, Leishman stain and Giemsa stain. All are used to ... Positive staining is more commonly used than negative staining in microbiology. The different types of positive staining are ... The dyes used in negative staining are acidic. Note: negative staining is a mild technique that may not destroy the ...
as a negative stain in biological electron microscopy, typically in the 3-5% (vol/vol) concentration range and in the presence ... "Negative staining", in Harris J. R. (Ed.), Electron Microscopy in Biology, Oxford University Press, Oxford. Adrian, Marc; ... De Carlo, S.; El-Bez, C.; Alvarez-Rúa, C.; Borge, J.; Dubochet, J. (2002). "Cryo-negative staining reduces electron-beam ... or at saturated concentration to perform cryo-negative staining. For the detection of recreational drugs as a component of the ...
When virions are coated with stain (positive staining), fine detail is obscured. Negative staining overcomes this problem by ... 33-37 Kiselev NA, Sherman MB, Tsuprun VL (1990). "Negative staining of proteins". Electron Microscopy Reviews. 3 (1): 43-72. ... Negative-sense viral RNA is complementary to mRNA and thus must be converted to positive-sense RNA by an RNA-dependent RNA ... To increase the contrast between viruses and the background, electron-dense "stains" are used. These are solutions of salts of ...
Gram negative, with bipolar staining. The presumptive identification of the pathogen is based on standard biochemical tests. In ... piscicida (previously known as Pasteurella piscicida) is a gram-negative rod-shaped bacterium that causes disease in fish. ...
Staining for actin, CD34 and smooth muscle actin are intermediate. Staining for S-100 protein is negative. Myxoid tumors ... hypocellular background Bland cytological appearance of spindle cells Immunohistochemical studies show strong staining for ...
Gram and Neisser staining reactions are negative. S. natans requires dissolved simple sugars or organic acids as a food supply ...
The diastase negative slide will show a magenta staining where glycogen is present within a section of tissue. The slide that ... A suitable basic stain is often used as a counterstain. • PAS diastase stain (PAS-D) is PAS stain used in combination with ... These cells stain a bright fuchsia. Pulmonary alveolar proteinosis. Fungal infection, the cell walls of fungi stain magenta; ... PAS staining is mainly used for staining structures containing a high proportion of carbohydrate macromolecules (glycogen, ...
Negative staining protocols typically treat the sample with 1% to 5% aqueous solution. Uranyl acetate staining is simple and ... Uranyl acetate is extensively used as a negative stain in electron microscopy. Most procedures in electron microscopy for ... doi:10.1016/0022-1902(75)80918-3. "Negative Staining" University of Oxford Sessler, Jonathan L.; Seidel, Daniel; Vivian, Anne E ... Some biological samples are not amenable to uranyl acetate staining and, in these cases, alternative staining techniques and or ...
Almeida, J. D.; Howatson, A. F. (1963). "A negative staining method for cell-associated virus". The Journal of Cell Biology. 16 ...
Almeida, J. D.; Howatson, A. F. (1963). "A negative staining method for cell-associated virus". The Journal of Cell Biology. 16 ... the first was a modified negative staining method using phosphotungstic acid, and the next was immunological procedure in which ... Several animals have been investigated and are proven to be negative. Among the possible carriers are Malayan pangolins (Manis ... whether the pathogen in B814 was a bacterium or a virus as all bacterial and viral culture methods available showed negative ...
... is a genus of Gram-negative staining bacteria. It includes the hydrocarbon-degrading strain Lutibacterium ...
Negative staining is also used for observation of nanoparticles.[citation needed] Sectioning - produces thin slices of the ... Negative stain - suspensions containing nanoparticles or fine biological material (such as viruses and bacteria) are briefly ... specimens can be stained "en bloc" before embedding and also later after sectioning. Typically thin sections are stained for ... Staining - uses heavy metals such as lead, uranium or tungsten to scatter imaging electrons and thus give contrast between ...
Virologists often use negative staining to help visualise viruses. In this procedure, the viruses are suspended in a solution ... Many viruses were discovered using this technique and negative staining electron microscopy is still a valuable weapon in a ... Brenner S, Horne RW (July 1959). "A negative staining method for high resolution electron microscopy of viruses". Biochimica et ... The separated molecules are revealed using stains such as coomasie blue, for proteins, or ethidium bromide for nucleic acids. ...
The background is stained while the organisms remain clear. This is called a negative stain. India ink, along with other stains ... Microbiologists use India ink to stain a slide containing micro-organisms. ...
... are negative by Gram stain. The order Anaeroplasmatales was created in 1987 to encompass the family ...
... permanent markers can be used to do a negative stain of a bacterial sample. This means the background is stained with the ... New version of the negative stain. Applied and Environmental Microbiology. 57:1858-1859. "Expo FAQs , Answers for Dry Erase ... The bacteria can be seen because they are unstained (lighter) while the background is stained (darker). Permanent markers are ... generally used on hard, non-porous surfaces, because instead of staining they form a surface layer that, despite their name, ...
Some organisms are gram-variable (meaning they may stain either negative or positive); some are not stained with either dye ... In addition, in all bacteria stained using the Gram stain, the age of the culture may influence the results of the stain. Gram- ... In microbiology and bacteriology, Gram stain (Gram staining or Gram's method), is a method of staining used to classify ... peptidoglycan and can stain negative, positive or indeterminate; members of the Deinococcota stain positive but are diderms ...
P. canis is a Gram-negative coccobacillus that shows bipolar staining. P. canis forms small, grey-colored, round, and smooth ... It can also be indole positive or negative depending on the biotype. Molecular analysis of the genome has become the popular ... Biberstein, E.L.; Jang, S.S.; Kass, P.H.; Hirsh, D.C. (1991). "Distribution of indole-producing urease-negative Pasteurellas in ... Before molecular technology was available, species were often labeled based on staining and the hosts from where species were ...
In 1959, he and Sydney Brenner pioneered the technique of negative staining in electron microscopy and used it to study virus ... BRENNER, S; HORNE, RW (July 1959). "A negative staining method for high resolution electron microscopy of viruses". Biochimica ... describing the development of negative staining (Articles with short description, Short description matches Wikidata, Articles ...
This is particularly useful in negative stain images. The algorithms make use of fast Fourier transforms (FFT), often employing ... Individual images of stained or unstained particles are very noisy, and so hard to interpret. Combining several digitized ... This low dose, as well as variations in the metal stain used (if used) means images have high noise relative to the signal ... Tilt methods are best suited to negatively stained samples, and can be used for particles that adsorb to the carbon support ...
Remaining cancer cells were weakly stained or negative. While interrogating 4440 tumor samples from 15 cancer types from TCGA, ... Nearby genes include SLC25A53 (on the negative strand) (about 8,000 base pairs (bps) upstream) and FAM199X (on the positive ... It is generally electroneutral (there are no positive or negative charge clusters or segments) with no high hydrophobic ...
Gram-negative bacteria are stained as pink colour. The chemical structure of the outer membrane's lipopolysaccharide is often ... of the cell wall in gram-negative bacteria. The gram-positive bacteria take up the crystal violet dye and are stained purple. ... Gram-negative cell walls are much thinner than the gram-positive cell walls, and they contain a second plasma membrane ... There are two main types of bacterial cell walls, those of gram-positive bacteria and those of gram-negative bacteria, which ...
The bacteria Gram stains negative and produce endospores. It was first "isolated from steamed Japanese cedar chips from a ...
Like all Proteobacteria the two species stain Gram-negative.(,cf.) They were isolated from coastal, surface waters of the north ...
They are ovoid in shape and stain Gram-negative. They are dependent on replication inside the host cells; thus, some species ... Gupta RS (August 2011). "Origin of diderm (Gram-negative) bacteria: antibiotic selection pressure rather than endosymbiosis ...
... and safranin using the gram staining procedure. Gram positive cells are purple after staining, while Gram negative cells are ... It is difficult to tell whether an organism is gram-positive or gram-negative using a microscope; gram staining, created by ... Coico R (October 2005). "Gram Staining". In Coico R, Kowalik T, Quarles J, Stevenson B (eds.). Current Protocols in ... The bacteria are stained with several dyes such as crystal violet, iodine alcohol, ...
It is a small, Gram-negative bacillus with bipolar staining by Wayson stain. In animals, it can originate in fulminant ...
... non-motile and stain Gram-negative. Strictly anaerobic organism. Chemo-organotrophic. Mucolytic in pure culture.: 1474 The ...
... myoD1 and myogenin stains yield negative test results. Thus, the detection of rhabdomyoblasts can play a key role in ... Staining sensitivity is highly variable from subtype to subtype. For instance the myogenin staining is far less efficient than ... The detection of benign nerve sheath tumors, a dense Schwannian cell background, negative test results for desmin, myogenin and ... These neoplastic cells yield positive results for vimentin, smooth muscle actin, and desmin stains; however, myoglobin, ...
"Negative staining for dummies". Retrieved 2009-06-06. "Negative staining". Archived from the original on 2012-12-25. Retrieved ... This contrasts with positive staining, in which the actual specimen is stained. For bright-field microscopy, negative staining ... The choice of negative stain in electron microscopy can be very important. An early study of plant viruses using negatively ... Negative staining is usually a very mild preparation method and thus does not reduce the possibility of operator infection. ...
Negative Staining Electron Microscope Protocol for Rash Illness ... Uranyl acetate (see Negative Stain Reagents section for recipe) ... Negative Staining Procedure. Negative Stain Reagents. *2% Phosphotungstic Acid (PTA): *2 g phosphotungstic acid in 100 mL dH20 ... Download: Negative Staining Electron Microscope Protocol for Rash Illness Cdc-pdf. [PDF - 10 pages] ... Stain. *Stains can be stored in 1 cc syringes equipped with syringe filters. Cover the syringe with 0.5% UA in foil to exclude ...
Negative staining: preparative procedures; Negative staining: some specialized approaches and problems; Negative staining: ... Negative Staining and Cryoelectron Microscopy The Thin Film Techniques By J.R. Harris Copyright 1996 ... Contents include : Introduction; Negative staining: historical background and technical development; ...
Negative staining is a procedure which embeds small biological molecules such as proteins, protein complexes, or nanoparticles ...
Antimicrobial activity against Pseudomonas aeruginosa assessed as cell damage at 150 uM after 60 mins by negative staining ...
A coated grid with sample adsorbed to the surface is floated on a drop of negative stain for 0.5-2 min, excess stain wicked ... Negative staining method for high resolution electron microscopy of viruses. Biochim Biophys Acta. 1959;34:103-10. DOIPubMed ... Negative Staining Particle Enrichment Identification of Viral Agents Future Impact of Diagnostic Electron Microscopy Cite This ... Negative staining in diagnostic virology. Micron and Micropica Acta. 1991;22:435-47. DOIGoogle Scholar ...
Negative staining of Asp23-strep. For transmission electron microscopy, Asp23-strep was subjected to negative staining to ... the negatives were dried at 40 °C overnight. Finally, the negatives were scanned (Quato Intelli Scan 1600, software Silverfast ... The exposed negatives were removed from the microscope, sorted to racks and transferred within the racks for 4 min into a box ... After two washing steps with water, the grids were stained with 1% aqueous uranyl acetate for 30 seconds. Air-dried grids were ...
... the observed negative results were concluded as the complete absence of staining or weak, discontinuous, and focal staining. ... P16 Immunostain Positive & Negative Meaning. P16 immunostaining can be both positive and negative. P16 or INK4A immunostain is ... However, the cytoplasmic staining as a whole was considered a negative one. ... Individuals or patients with HPV16 negative or P16 negative status are known; to show the worst survival rate for all of the ...
Empiric therapy should be guided by results of synovial fluid Gram staining. Initial therapy should consist of IV antibiotics. ... The Gram stain is positive in less than 50% of cases. When the Gram stain is negative and there are no crystals apparent, it ... Gram stain negative or unavailable Diagnosis of the septic joint is particularly challenging in patients with underlying ... What is the workup and treatment of Gram stain negative septic arthritis of native joints? ...
Empiric therapy should be guided by results of synovial fluid Gram staining. Initial therapy should consist of IV antibiotics. ... The Gram stain is positive in less than 50% of cases. When the Gram stain is negative and there are no crystals apparent, it ... Gram stain negative or unavailable Diagnosis of the septic joint is particularly challenging in patients with underlying ... What is the workup and treatment of Gram stain negative septic arthritis of native joints? ...
1). The negative-stain EM images showed that each extracted oligomer of YaxAB-C8, YaxAB-C9, or YaxAB-C10 has 8, 9, or 10 ... Negative-stain electron microscopy (EM) and fitting model structure. BN-PAGE-extracted YaxAB-C8 pores were applied to glow- ... Raw micrographs of negative-staining electron microscopy (EM) for pores generated in this study have been deposited in the ... The initial model of YaxAB-C8 for the negative stain 3D reconstruction was generated from three representative classes of 2D ...
Gram Negative bacilli, Gram negative cocci, Gram Stain, Gram-Negative Bacilli and Coccobacilli Bacteria, Gram-Negative Bacilli ... Gram-Negative Coccobacilli of Kingella kingae in Gram staining of Culture Gram-Negative Bacilli and Coccobacilli Bacteria: ... Introduction of Gram-Negative Bacilli and Coccobacilli Bacteria Gram-Positive Negative (GNB) is also called Gram-Negative ... Gram-Negative Coccobacilli of Kingella kingae in Gram staining of Sputum, Gram-Negative Coccobacilli of Kingella kingae in Gram ...
The patients in our study were diagnosed with triple-negative breast cancer in Singapore General Hospital from 2003 to 2015. We ... staining or Massons trichrome (MT) staining. However, this analysis is highly dependent on the staining protocol and color ... Quantitative stain-free imaging and digital profiling of collagen structure reveal diverse survival of triple negative breast ... Quantitative stain-free imaging and digital profiling of collagen structure reveal diverse survival of triple negative breast ...
EM; negative staining (3); thin-section (20) Surface Projections/Envelope. Spiked virions (20); spherical 7 mm capsomere-like ...
Negative-stain electron microscopy. Negative stain specimens were prepared by applying 4 µL of protein solution (0.1 mg/mL + 1 ... E) Representative negative stain transmission electron micrographs showing that purified 6XHis-PilTK136A and 6XHis-PilUK134A ... A) Representative negative stain transmission electron micrographs of purified 6XHis-PilT and 6XHis-PilU (left; scale bar, 50 ... Negative stain transmission electron microscopy revealed that both 6XHis-PilT and 6XHis-PilU formed hexameric ring complexes ( ...
Negative staining. Request a detailed protocol Overnight cultures were fixed in 2% glutaraldehyde and negatively-stained with 2 ... Right: Analysis of clinical isolates using a negative-staining to identify the presence of flagella. (E) Phagocytic uptake of ... As demonstrated by electron microscopy with negative-staining the motile isolate was flagellated, while one of the non-motile ... Negative Stain.. https://cdn.elifesciences.org/articles/55744/elife-55744-fig2-data1-v2.pdf Download elife-55744-fig2-data1-v2. ...
Combined with a high-throughput procedure for preparation of EM-grids and automation of the subsequent negative staining step, ...
Is a cDNA footprint signal in my Tissue Optimization experiment negative control with IF staining expected? Answer: The... ... Is a cDNA footprint signal in my Tissue Optimization experiment negative control with IF staining expected? Question: Is a cDNA ... Observing low fluorescence is expected with immunofluorescence (IF) stained tissue sections. The blocking, staining, and ... Answer: The negative control during the Tissue Optimization (TO) experiment may show minimal fluorescence signal. It serves as ...
Negative staining proteins and macromolecules. *Nanoparticle analysis. *Tomography. *Immunogold labelling. *CLEM. Zeiss Gemini ...
... negative staining, whole-mount SEM, correlative LM and EM, and Immunogold labeling. ... User negative staining. $5 plus $5 per glow discharge run. Cost of materials (grids extra). ... Sectioning & staining. $65 per hour. Typically 1 hour per block. SEM processing. $55 per sample. A $20 per run surcharge will ...
A) Negative staining electronic microscopy observation. (B-L) Transmission electronic microscopy images. (A) Morphology of ... A) Negative staining electronic microscopy observation. (B-L) Transmission electronic microscopy images. (A) Morphology of ... In addition, the negative effect of virophages is different from that observed with some satellite viruses as for the case of ... The negative effect of virophages on their viral hosts is probably the strongest support of the virophage concept. However, it ...
Negative-stain transmission electron microscopy. Request a detailed protocol Wild-type S. cerevisiae 26S holoenzyme was diluted ... 11) Since the negative-stain EM analysis is the major evidence for the Rpn5-VTENKIF mutant perturbing the natural distribution ... 11) Since the negative-stain EM analysis is the major evidence for the Rpn5-VTENKIF mutant perturbing the natural distribution ... We employed negative-stain electron microscopy to assess the conformational states of the proteasome and whether the Rpn5- ...
Herpesvirus (negative staining) cc-by-sa-3.0. Dr Graham Beards Wikimedia Commons ... Living cells were stained with crystal violet. The viral plaques, each was from one virion, remained transparent. Date: 1 March ... This negatively-stained transmission electron micrograph (TEM) revealed the presence of numerous herpes simplex type-2 virions ... This negatively-stained transmission electron micrograph (TEM) revealed the presence of numerous herpes simplex virions, ...
MRK16 staining intensity is categorized as follows: negative (d , 0.10), dim (0.10 , D , 0.15), moderate (0.15 , D , 0.25) and ... with surface staining and/or DNA staining optional *Optional, for surface staining: Stain cells using standard surface staining ... with surface staining and/or DNA staining optional *Optional: For surface marker staining, stain cells with desired surface ... 13.2 Cell Surface Staining (Ab mix from the bank is for a final staining volume of 50uL) *Note the volume of Ab mix per sample ...
No positive staining was observed in the negative control sections (Alexa Fluor 488 alone). Scale bars: 100 μm; original ... No positive staining was observed in the negative control sections. Scale bars: 100 μm; original magnification, ×60 and ×120 ( ... Ad-p27 and Ad-p27-126TS inhibit restenosis 2 weeks after balloon injury. (A) Representative H&E-stained sections. Scale bars: ... These sections were negative for GFP expression (Supplemental Figure 2A, uninjured). Moreover, we verified p27 overexpression ...
It is a negative staining method - background is stained not the capsule • Sensitive test for AIDS patients (90% sensitive) • ... Positive [red] on the Modified Kinyoun acid fast stain. Modified acid-fast stain [modified Kinyoun stain] Gram stain of sputum ... Grocotts Methenamine Silver Stain [GMS] - yeast and hyphae stain grey to black. Will stain living and dead fungi • Observe the ... "negative" stain - which stains the background not the yeast cell. The clearing around the yeast is due to the polysaccharide ...
The negative staining controls omitted the primary antibodies. Under these conditions, staining was completely abolished. ... The blue staining represents nuclei stained by DAPI. The yellow merged images show positive co-localization of TNF-R1 in ... Blue staining represents fluorescent nuclei stained using DAPI. (B) Quantification of the fluorescence of pinealocytes ... Glial cells are stained in red: astrocytes [GFAP-labeled, (B)] and microglia [ED-1-labeled, (E)]. GFAP-positive cells are ...
Categories: Negative Staining Image Types: Photo, Illustrations, Video, Color, Black&White, PublicDomain, CopyrightRestricted 1 ...
LC3 staining was negative in airway epithelial cells (Supplemental Figure 2B). Higher levels of LC3II in isolated AECIIs from ... PINK1 deficiency increases susceptibility to lung fibrosis. (A) Representative Masson trichrome staining in Pink1+/+, Pink1+/- ... LC3 staining was higher in hyperplasic AECIIs from fibrotic areas in IPF lungs (Figure 3A and Supplemental Figure 2A). Analyses ... Epithelial cells from honeycomb areas (B and C) showed positive staining for both markers. Epithelial cells from areas with ...
The effectiveness of the proposed system was tested over 21 histologically stained images containing approximately 1800 cell ... an automated entropy-based thresholding system for segmentation and quantification of cell nuclei from histologically stained ... Positive and negative control sections were included to ensure reproducible staining. Hematoxylin & Eosin (H&E) stain was used ... stain, whereas the proposed entropy-based thresholding algorithm segments cell nuclei from images stained by H&E and three more ...
  • Type IV pili are ubiquitous surface appendages in Gram-negative bacteria that promote diverse activities including attachment, virulence, biofilm formation, horizontal gene transfer, and twitching motility [ 1 - 5 ]. (biorxiv.org)
  • When we fully understand the structure and function of TonB, ExbB, and ExbD, then we will know a critical mechanism whereby Gram-negative bacteria acquire iron. (mcgill.ca)
  • Why are gram-positive bacteria typicaly more resistant than gram-negative bacteria to antibiotics, that disrupt plasma membranes, such as polymyxin B? (answerbag.com)
  • Danish bacteriologist who in 1884 developed a method of staining bacteria, called Gram's stain or Gram's dye, that is used to identify and classify bacteria, often from samples of infected body fluids. (dictionary.com)
  • A Gram stain is a test that checks for bacteria at the site of a suspected infection such as the throat, lungs, genitals, or in skin wounds. (medlineplus.gov)
  • Gram stains may also be used to check for bacteria in certain body fluids, such as blood or urine. (medlineplus.gov)
  • The categories are diagnosed based on the how the bacteria reacts to the Gram stain. (medlineplus.gov)
  • When the stain combines with bacteria in a sample, the bacteria will either stay purple or turn pink or red. (medlineplus.gov)
  • If the bacteria turns pink or red, they are Gram-negative. (medlineplus.gov)
  • Knowing whether bacteria is Gram-positive or Gram-negative can help your provider identify the type of infection you have and which antibiotics will be most effective in treating it. (medlineplus.gov)
  • If the bacteria was colored pink or red, it means you likely have a Gram-negative infection. (medlineplus.gov)
  • Gram-negative bacteria are classified by the color they turn after a chemical process called Gram staining is used on them. (msdmanuals.com)
  • Gram-negative bacteria stain red when this process is used. (msdmanuals.com)
  • Gram-positive bacteria stain blue. (msdmanuals.com)
  • Gram-negative and gram-positive bacteria stain differently because their cell walls are different. (msdmanuals.com)
  • Gram-positive bacteria are classified by the color they turn after a chemical called Gram stain is applied to them. (msdmanuals.com)
  • Gram-positive bacteria stain blue when this stain is applied to them. (msdmanuals.com)
  • Other bacteria stain red. (msdmanuals.com)
  • Infections caused by bacteria that show up as pink (negative) when treated by the gram-staining method. (bvsalud.org)
  • The choice of negative stain in electron microscopy can be very important. (wikipedia.org)
  • Negative staining at both light microscope and electron microscope level should never be performed with infectious organisms unless stringent safety precautions are followed. (wikipedia.org)
  • Negative staining transmission electron microscopy has also been successfully employed for study and identification of aqueous lipid aggregates like lamellar liposomes (le), inverted spherical micelles (M) and inverted hexagonal HII cylindrical (H) phases (see figure). (wikipedia.org)
  • Electron microscopic (EM) visualization of negatively stained poxvirus virions was a valuable technique for confirming poxvirus infections during the smallpox eradication campaign. (cdc.gov)
  • Unless you have electron-dense nanoparticles (such as gold), a heavy metal stain is necessary to visualize the relatively electron-translucent materials. (ki.se)
  • Variations on Negative Stain Electron Microscopy Methods: Tools for Tackling Challenging Systems. (ki.se)
  • Negative and Positive Staining in Transmission Electron Microscopy for Virus Diagnosis. (ki.se)
  • After a simple and fast negative stain preparation, the undirected, "open view" of electron microscopy allows rapid morphologic identification and differential diagnosis of different agents contained in the specimen. (cdc.gov)
  • Combined with a high-throughput procedure for preparation of EM-grids and automation of the subsequent negative staining step, the crystallization block offers a novel pipeline that promises to speed up large-scale screening of 2D crystallization and to increase the likelihood of producing well-ordered crystals for analysis by electron crystallography. (nih.gov)
  • This negatively-stained transmission electron micrograph (TEM) revealed the presence of numerous herpes simplex virions, located inside a cell nucleus in this tissue sample. (eol.org)
  • This negatively-stained transmission electron micrograph (TEM) revealed the presence of numerous herpes simplex type-2 virions, members of the Herpesviridae virus family. (eol.org)
  • Medium resolution electron microscopy resolved particles (10 nm diameter) by negative staining (Sverzhinsky et al. (mcgill.ca)
  • In microscopy, negative staining is an established method, often used in diagnostic microscopy, for contrasting a thin specimen with an optically opaque fluid. (wikipedia.org)
  • For bright-field microscopy, negative staining is typically performed using a black ink fluid such as nigrosin and India ink. (wikipedia.org)
  • Gram stains, cultures, and nucleic acid amplification tests (NAATs) of genital, rectal, conjunctival, pharyngeal secretions and/or other extragenital sites, as applicable, should also be obtained when DGI is suspected. (medscape.com)
  • This term earlier described immunohistochemical staining of tissue sections by Albert Coons in 1941. (healthncare.info)
  • This term was first described by Albert Coons to refer to the immunohistochemical staining of tissue sections. (healthncare.info)
  • Immunohistochemical staining showed that they were negative for CD68, S-100, vimentin, and HMB-45. (karger.com)
  • In the event of a deliberate release of smallpox virus and subsequent human disease, or in generalized vaccinia infections resulting from vaccination, negatively stained preparations derived from lesions or scab material would again provide a valuable method for assisting in poxvirus diagnosis and/or ruling out other causes of rash illness. (cdc.gov)
  • Diagnosis of the septic joint is particularly challenging in patients with underlying inflammatory disease, such as rheumatoid arthritis or SLE, and a negative Gram stain. (medscape.com)
  • In this technique, the background is stained, leaving the actual specimen untouched, and thus visible. (wikipedia.org)
  • This contrasts with positive staining, in which the actual specimen is stained. (wikipedia.org)
  • The specimen, such as a wet bacterial culture spread on a glass slide, is mixed with the negative stain and allowed to dry. (wikipedia.org)
  • Gram stain - Most specimen types can be Gram stained. (slideshare.net)
  • The negative electrons rush toward it and those which are in the centre fly, accelerated, through the orifice toward the specimen. (magma.ca)
  • Negative staining is a procedure which embeds small biological molecules such as proteins, protein complexes, or nanoparticles on a TEM grid in a thin film of heavy metal salts (i.e. uranyl salts) to enhance contrast and reveal their structural details. (ki.se)
  • Visualizing proteins and macromolecular complexes by negative stain EM: from grid preparation to image acquisition. (ki.se)
  • This is demonstrated in the TO experiment images shown below where 10 µm tissue sections from mouse brain were stained with DAPI (blue) using the Methanol Fixation, Immunofluorescence Staining & Imaging Demonstrated Protocol. (10xgenomics.com)
  • However, they showed negative coloration after G- and restriction enzyme banding and DAPI staining. (ingentaconnect.com)
  • Observing low fluorescence is expected with immunofluorescence (IF) stained tissue sections. (10xgenomics.com)
  • The primary tasks involved in the analysis of histologically stained tissue sections are cell nuclei counting, detecting abnormal cell nuclei, and the presence of antigens within the target cells. (hindawi.com)
  • The blocking, staining, and washing buffers used in the IF protocol contain a small amount of surfactant that enables removal of non-specific antibody binding as well as permeabilization of the tissue in order to facilitate intracellular protein staining. (10xgenomics.com)
  • Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. (abcam.com)
  • Traditionally, uranyl acetate or uranyl formate are used for staining followed by room temperature TEM (Talos). (ki.se)
  • The structures which can be negatively stained are much smaller than those studied with the light microscope. (wikipedia.org)
  • An early study of plant viruses using negatively stained leaf dips from a diseased plant showed only spherical viruses with one stain and only rod-shaped viruses with another. (wikipedia.org)
  • For EphA2 expression, 21 tumors (10%) showed negative, 108 (50%) weak positive, 69 (32%) moderate positive and 19 (9%) strong positive, whereas for EphrinA-1 expression, 33 tumors (15%) showed negative, 91 (42%) weak positive, 67 (31%) moderate positive and 26 (12%) strong positive. (medsci.org)
  • Interestingly, tumors with both negative mdm-2 and p21 expression, irrespective of p53 status, had a high response rate to docetaxel but no response to MF. (lu.se)
  • The factors responsible for response to tamoxifen in 5% to 10% of patients with ERa-negative tumors are not clear. (lu.se)
  • negative tumors do seem to be sensitive to tamoxifen (5 - 7). (lu.se)
  • In univariate analysis high expression (strong staining) of EphrinA-1 was associated with poor disease-free ( P = 0.033) and disease-specific ( P = 0.039) survival. (medsci.org)
  • Historically, negative stain EM successfully detected poxvirus particles in approximately 95% of clinical specimens from patients with orthopoxvirus infections such as variola (smallpox)/monkeypox, and approximately 65% from patients with vaccinia (smallpox vaccine). (cdc.gov)
  • Negative staining for biological particles is a standard EM technique that allows you to investigate the size and structure of biomolecules and nanoparticles, achieving resolutions up to 20 Å. (ki.se)
  • In this process, particles are surrounded by the stain, which scatters electrons strongly creating negative contrast, whereby the particle appears bright against the stain. (ki.se)
  • Parameters such as staining time and blotting technique affect the proportions between negative and positive staining on the grid and may need to be optimized for each sample. (ki.se)
  • As strong oxidants, they cross-link lipids mainly by reacting with unsaturated carbon-carbon bonds, and thereby both fix biological membranes in place in tissue samples and simultaneously stain them. (wikipedia.org)
  • Recognition of the pneumococcus depends upon its morphology, the fact that it is gram -staining, and the presence of a capsule. (dictionary.com)
  • You should not co-stain in IHC antigens located on the same structure (nucleus, membrane, cytoplasm). (ihcworld.com)
  • Results of IHC staining of skin, fat, muscle, and bone marrow samples were negative for West Nile Virus antigens. (cdc.gov)
  • EM laboratories involved in negative stain viral diagnostics are encouraged to participate in the External Quality Assessment program External administered by the Robert Koch Institut in Berlin. (cdc.gov)
  • Negative staining is usually a very mild preparation method and thus does not reduce the possibility of operator infection. (wikipedia.org)
  • When the Gram stain is negative and there are no crystals apparent, it may be reasonable to withhold antibiotics and treat for a crystalline arthritis, unless there is a significant potential source of bacteremia such as a urinary tract infection or the patient exhibits significant systemic deterioration. (medscape.com)
  • A Gram stain is most often used to find out if you have a bacterial infection. (medlineplus.gov)
  • If you do, the test will show if your infection is Gram-positive or Gram-negative. (medlineplus.gov)
  • Gram stain results may also show whether you have a fungal infection. (medlineplus.gov)
  • A cell nuclei segmentation algorithm incorporating unsupervised color clustering, morphological operations, and local thresholding has been proposed to distinguish the cancerous and noncancerous areas in histologically stained images and then segment the clustered cell nuclei [ 3 ]. (hindawi.com)
  • To identify the gram-negative rods grown from blood cultures and a right-knee fluid aspirate from an 80-year-old caucasian man who had undergone a total right knee arthroplastic procedure 6 years ago, and to assess the genetic similarity between the 2 isolates. (medscape.com)
  • Two months prior to her initial presentation to us she was prescribed 6 months of 4-drug TB therapy because of the hemoptysis and an abnormal chest x-ray [Image A] even though she had 3 negative sputa for acid-fast bacilli. (uab.edu)
  • [Image G] . Paragonimus eggs are destroyed in sputum being processed for acid-fast staining so sputum needs to be processed for O&P detection in order to detect eggs. (uab.edu)
  • Acid fast stain was negative. (uab.edu)
  • There are granulomas in the reticular dermis ( Image E ). Fite-Faraco stain is 2+ positive ( Image F ). In order to demonstrate acid-fast bacilli in tissue, a Fite-Faraco stain is necessary as a Ziehl-Neelsen stain may be negative. (uab.edu)
  • How do acid-fast properties relate to the gram stain? (answerbag.com)
  • However, the observed negative results were concluded as the complete absence of staining or weak, discontinuous, and focal staining. (healthncare.info)
  • 2 methods: histopathological examination tion is focal, negative biopsy results do not and bacterial culture growth. (who.int)
  • First to consider is the diagnostic capabilities of the laboratory personnel, who should have experience with preparing negative stain EM grids, and, more importantly, must have experience with analyzing negative stain EM preparations to identify virus morphology, and to differentiate these from look-alikes and artifacts. (cdc.gov)
  • You don't need any special preparations for a Gram stain. (medlineplus.gov)
  • Examine stain for budding cells to confirm that it is a yeast cell and not an artifact. (slideshare.net)
  • Negative staining is often used in conjunction with cryo-EM to check the sample before freezing, to ensure that the protein complex is intact, the sample is not aggregated, and the concentration is appropriate. (ki.se)
  • • Yeast cells stain blue [Gram positive]. (slideshare.net)
  • Image 4: GMS staining better defines individual yeast making recognition easier at scanning magnifications. (cap.org)
  • Results: It was observed that the pulp stem cells exhibited the capacity to adhere to plastic and a high rate of expansion and, after detection with specific stains, it was shown that the cells were capable of differentiation into osteoblasts and chondroblasts, but not into adipocytes. (bvsalud.org)
  • The PP16 or HPV16 immunostain negative also means that it is more likely a distinct subgroup that lacks any HPV-related genotype. (healthncare.info)
  • P = 0.01) within the ERa- negative subgroup even after adjustment for other markers. (lu.se)
  • The cytoplasm itself was negative on periodic acid Schiff (PAS) and Sudan staining. (karger.com)
  • That because the success of this double staining procedure is based on the complete development of the first stain, which should mask the structure stained first. (ihcworld.com)
  • Careful check possible crossreactivty of your secondary antibodies by staining in simple IHC the relevant heterologous primary antibodies. (ihcworld.com)
  • The donor's serum sample was positive for West Nile Virus IgM, IgG, and neutralizing antibodies by serologic testing but negative for West Nile Virus RNA by nucleic acid amplification testing. (cdc.gov)
  • Purpose: Endocrine therapies, such as tamoxifen, are commonly given to most patients with estrogen receptor (ERa) ^ positive breast carcinoma but are not indicated for persons with ERa-negative cancer. (lu.se)
  • These results may be highly clinically significant, because in the United States alone, f10,000 women are diagnosed annually with ERa-negative/ERh-positive breast carcinoma and may benefit from adjuvant tamoxifen. (lu.se)
  • Analysis of the cellular phenotype showed that the cells were negative for CD45, CD69, CD117 and HLADR, and positive for CD13, CD44, CD73, CD90 and CD105. (bvsalud.org)
  • In patients with negative P16, things are usually more complicated than the patients exhibiting positive P16 immunostaining results. (healthncare.info)
  • An alternative method has been developed using an ordinary waterproof marking pen to deliver the negative stain. (wikipedia.org)
  • The great majority belong to the colon bacillus group, and are negative to gram 's method of staining. (dictionary.com)
  • Empiric antibiotic therapy of septic arthritis of native joints(SA) has been guided primarily by the findings of the Gram staining of synovial fluid. (medscape.com)
  • The most common types of Gram stain tests are listed below. (medlineplus.gov)
  • It serves as a baseline for determining "background" fluorescence, allowing customers to reduce the laser power or gain such that the negative control fluorescence is no longer observed. (10xgenomics.com)
  • 12. Check the staining until complete, dense staining is obtained, but background is still low. (ihcworld.com)
  • P16 immunostaining can be both positive and negative. (healthncare.info)
  • 11. After 5 min, check the staining in your positive and negative controls. (ihcworld.com)
  • Therefore, it is important to make individualized treatment procedures in order to reduce negative side effects for patients with a good prognosis. (medsci.org)
  • Negative staining can also be used independently from cryo-EM to determine a 3D structure. (ki.se)