'Recent advances in gold technology have led to probes with improved properties and performance for cell biologists: higher labeling density, better sensitivity, and greater penetration into tissues. Gold clusters, such as the 1.4-nm Nanogold, are gold compounds that can be covalently linked to Fab'' antibody fragments, making small and stable ... More
Use of nanogold- and fluorescent-labeled antibody Fv fragments in immunocytochemistry.
AuthorsRibrioux S, Kleymann G, Haase W, Heitmann K, Ostermeier C, Michel H
JournalJ Histochem Cytochem
PubMed ID8648079
'Recombinant antibody fragments are emerging as a versatile tool in both basic research and medical therapy. We describe the procedures for direct labeling of engineered antibody fragments (Fv) with fluorescein or nanogold and their use in fluorescence and immunoelectron microscopy, respectively. The Fv fragments were produced in Escherichia coli, purified ... More
Cryo-electron microscopic localization of protein L7/L12 within the Escherichia coli 70 S ribosome by difference mapping and Nanogold labeling.
AuthorsMontesano-Roditis L, Glitz DG, Traut RR, Stewart PL
JournalJ Biol Chem
PubMed ID11278411
'The Escherichia coli ribosomal protein L7/L12 is central to the translocation step of translation, and it is known to be flexible under some conditions. The assignment of electron density to L7/L12 was not possible in the recent 2.4 A resolution x-ray crystallographic structure (Ban, N., Nissen, P., Hansen, J., Moore, ... More
DNA monolayer on gold substrates characterized by nanoparticle labeling and scanning force microscopy.
AuthorsCsáki A, Möller R, Straube W, Köhler JM, Fritzsche W
JournalNucleic Acids Res
PubMed ID11504889
'Monolayers of single-stranded DNA on gold substrates were studied by scanning force microscopy. Complementary DNA probes labeled by gold nanoparticles were applied for contrast enhancement. Substrate regions modified with DNA could be visualized in a highly specific manner. The influence of the solution concentration on the surface density of adsorbed ... More
Concomitant oncoprotein detection with fluorescence in situ hybridization (CODFISH): a fluorescence-based assay enabling simultaneous visualization of gene amplification and encoded protein expression.
'We sought the validation of a three-color fluorescence-based system that simultaneously profiles Her2/neu oncogene copy by fluorescence in situ hybridization (FISH) and Her-2/neu encoded protein by the use of a versatile alkaline phosphatase chromogen fast red K in either fluorescence or bright-field mode. Nuclei were counterstained with DAPI. Nineteen infiltrating ... More
Labeling with nanogold and undecagold: techniques and results.
AuthorsHainfeld JF
JournalScanning Microsc Suppl
PubMed ID9601549
A significant new development in gold labeling for microscopy has been achieved through the use of gold cluster compounds that are covalently attached to antibodies or other probe molecules. These unique gold probes are smaller than most colloidal gold conjugates and exhibit improved penetration into tissues, higher labeling densities, and ... More
"Plugging into Enzymes": nanowiring of redox enzymes by a gold nanoparticle.
AuthorsXiao Y, Patolsky F, Katz E, Hainfeld JF, Willner I
JournalScience
PubMed ID12649477
The reconstitution of an apo-flavoenzyme, apo-glucose oxidase, on a 1.4-nanometer gold nanocrystal functionalized with the cofactor flavin adenine dinucleotide and integrated into a conductive film yields a bioelectrocatalytic system with exceptional electrical contact with the electrode support. The electron transfer turnover rate of the reconstituted bioelectrocatalyst is approximately 5000 per ... More
Immunogold labelling of neuroendocrine peptides with special reference to antibody specificity and multiple staining techniques.
AuthorsLarsson LI
JournalHistochem Cell Biol
PubMed ID8858369
Immunogold methods have been very important for research on the neuroendocrine system. The compatibility of immunogold probes with optimal contrasting for electron microscopy has made localizations of neuroendocrine peptides to different subtypes of secretory organelles possible and, currently, methods using covalent attachment of nanogold particles to antibodies and neuropeptide ligands ... More
Combined fluorescent and gold immunoprobes: reagents and methods for correlative light and electron microscopy.
AuthorsPowell RD, Halsey CM, Hainfeld JF
JournalMicrosc Res Tech
PubMed ID9712158
Immunoprobes which incorporate both a fluorescent label and a 1.4 nm gold cluster compound were prepared by covalent conjugation to Fab' antibody fragments of the Nanogold cluster label followed by a fluorescent moiety. These new immunoconjugates allow the collection of two complementary sets of data, from fluorescence and electron microscopy, ... More
Remote electronic control of DNA hybridization through inductive coupling to an attached metal nanocrystal antenna.
AuthorsHamad-Schifferli K, Schwartz JJ, Santos AT, Zhang S, Jacobson JM
JournalNature
PubMed ID11805829
Increasingly detailed structural and dynamic studies are highlighting the precision with which biomolecules execute often complex tasks at the molecular scale. The efficiency and versatility of these processes have inspired many attempts to mimic or harness them. To date, biomolecules have been used to perform computational operations and actuation, to ... More
Lipopolysaccharide internalization activates endotoxin-dependent signal transduction in cardiomyocytes.
AuthorsCowan DB, Noria S, Stamm C, Garcia LM, Poutias DN, del Nido PJ, McGowan FX
JournalCirc Res
PubMed ID11249872
We tested the hypothesis that bacterial lipopolysaccharide (LPS) must be internalized to facilitate endotoxin-dependent signal activation in cardiac myocytes. Fluorescently labeled LPS was used to treat primary cardiomyocyte cultures, perfused heart preparations, and the RAW264.7 macrophage cell line. Using confocal microscopy and spectrofluorometry, we found that LPS was rapidly internalized ... More
Direct visualization of the calmodulin subunit of phosphorylase kinase via electron microscopy following subunit exchange.
Calmodulin is a tightly bound, intrinsic subunit (delta) of the hexadecameric phosphorylase-b kinase holoenzyme, (alphabetagammadelta)4. To introduce specifically labeled calmodulin into the phosphorylase-b kinase complex for its eventual visualization by electron microscopy, we have developed a method for rapidly exchanging exogenous calmodulin for the intrinsic delta subunit. This method exploits ... More