Search results for "Negative stain"
showing 7 items of 37 documents
Electron microscopy and biochemical characterization of a 350-kDa annular hemolymph protein from the keyhole limpet Megathura crenulata
1994
The isolation and biochemical characterization of an annular non-hemocyanin hemolymph protein from a marine gastropod, the Californian giant keyhole limpet (Megathura crenulata) is presented. By analytical ultracentrifugation, the protein has a sedimentation coefficient of 12S and molecular mass of approximately 350 kDa. The subunit mass, obtained by SDS/PAGE in the presence of -SH reagent and 8 M urea, is approximately 35 kDa, thereby indicating the presence of 10 subunits in the native molecule. By negative staining, the protein is revealed in one predominant image projection as a pentagonal approximately 8 nm ring-like structure with an approximately 2-nm stain-filled centre and, in anot…
Cryptosporidium parvum: Structural Components of the Oocyst Wall
1999
Cryptosporidium parvum, an enteropathogenic parasite, infects a wide range of mammals including man and constitutes a substantial veterinary and medical threat due to its ubiquitous distribution and the stability of the oocyst stage. The oocyst wall of C. parvum is known to be extremely resistant to chemical and mechanical disruption. Isolated oocyst walls are shown by both thin sectioning and negative staining transmission electron microscopy to possess a filamentous array on the inner surface. This filamentous array can be greatly depleted by digestion with proteinase K and trypsin, but pepsin has less effect. Ultrasonication of the untreated oocyst walls produced almost no fragmentation,…
Immunoelectron Microscopy of Hemocyanin from the Keyhole Limpet (Megathura crenulata): A Parallel Subunit Model
1993
Abstract Immunoelectron microscopy has been performed using negatively stained immune complexes of keyhole limpet hemocyanin (KLH) subunit 2 di- and multidecamers with domain-specific monoclonal antibodies. One antibody (KLH2 a macr 1) links the hemocyanin molecules in a side-to-side pattern, whereas the other antibody (KLH2 fg macr 1) links the molecules end-to-end. From existing knowledge of the domain sequence of KLH subunit 2, these data provide support for a parallel arrangement of subunits within each decamer. Ten N-terminal a macr: domains are then present at the noncollar region of each decamer with 10 C-terminal g macr domains at the collar region. The immunonegative staining data …
Application of the negative staining technique to both aqueous and organic solvent solutions of polymer particles
1999
Abstract Transmission electron microscopy (TEM) imaging of several different polymers in aqueous and organic solutions using the negative staining technique is demonstrated, to emphasise the possibilities of this specimen preparation technique for polymer science. Negative stains can readily be prepared in both water and organic solvents (e.g. dimethyl formamide (DMF), dimethyl sulfoxide (DMSO) and tetrahydrofurane (THF)). Polymer particle size, size distribution and shape seen in negative stain correlates well with those of unstained materials. The particle surface and smaller particles (e.g. 10–20 nm) are more clearly defined in the presence of a negative stain. The inherent problems of s…
Negative Staining of Thinly Spread Biological Samples
2007
Negative staining is widely applicable to isolated viruses, protein molecules, macro-molecular assemblies and fibrils, subcellular membrane fractions, liposomes and artificial membranes, synthetic DNA arrays, and also to polymer solutions. In this chapter, techniques are provided for the preparation of the necessary support films (continuous carbon and holey/perforated carbon). The range of suitable negative stains is presented, with some emphasis on the benefit of using ammonium molybdate and of negative stain-trehalose combinations. Protocols are provided for the single-droplet negative staining technique (on continuous and holey carbon support films), the negative staining-carbon film te…
The structure of gas-filled n-butyl-2-cyanoacrylate (BCA) polymer particles
1995
Abstract The structure of gas-filled poly-[n-butyl-2-cyanoacrylate] (BCA) particles has been demonstrated by negative staining with uranyl acetate, platinum-carbon shadowing of air-dried material and thin sectioning of the aqueous suspension of BCA particles, embedded in water-soluble melamine resin. The polymer shell of the hollow particles possesses a globular outer surface and a smoother inner surface.
Haliotis tuberculata hemocyanin (HtH): analysis of oligomeric stability of HtH1 and HtH2, and comparison with keyhole limpet hemocyanin KLH1 and KLH2
2000
The multimeric/higher oligomeric states of the two isoforms of Haliotis tuberculata hemocyanin (HtH1 and HtH2) have been assessed by transmission electron microscopy (TEM) of negatively stained specimens, for comparison with previously published structural data from keyhole limpet hemocyanin (KLH1 and KLH2) [see Harris, J.R., Gebauer, W., Guderian, F.U., Markl, J., 1997a. Keyhole limpet hemocyanin (KLH), I: Reassociation from Immucothel followed by separation of KLH1 and KLH2. Micron, 28, 31-41; Harris, J.R., Gebauer, W., Söhngen, S.M., Nermut, M.V., Markl, J., 1997b. Keyhole limpet hemocyanin (KLH). II: Characteristic reassociation properties of purified KLH1 and KLH2. Micron, 28, 43-56; H…