The ORF YOL018c (TLG2) of Saccharomyces cerevisiae encodes a protein that belongs to the syntaxin protein family. The proteins of this family, t-SNAREs, are present on target organelles and are thought to participate in the specific interaction between vesicles and acceptor membranes in intracellular membrane trafficking. TLG2 is not an ... More
Increased number of synaptic GABA(A) receptors underlies potentiation at hippocampal inhibitory synapses.
AuthorsNusser Z, Hájos N, Somogyi P, Mody I
JournalNature
PubMed ID9744275
Changes in synaptic efficacy are essential for neuronal development, learning and memory formation and for pathological states of neuronal excitability, including temporal-lobe epilepsy. At synapses, where there is a high probability of opening of postsynaptic receptors, all of which are occupied by the released transmitter, the most effective means of ... More
Sensitive in situ hybridization with catalyzed reporter deposition, streptavidin-Nanogold, and silver acetate autometallography: detection of single-copy human papillomavirus.
AuthorsZehbe I, Hacker GW, Su H, Hauser-Kronberger C, Hainfeld JF, Tubbs R
JournalAm J Pathol
PubMed ID9137082
The usefulness of standard in situ hybridization for viral nucleic acid detection is occasionally limited by its sensitivity limit of 10 to 50 copies per cell. A modified version of the recently described signal amplification method, catalyzed reporter deposition (CARD), and its application to formalin-fixed cells and tissue sections is ... More
Morphology of the yeast endocytic pathway.
AuthorsPrescianotto-Baschong C, Riezman H
JournalMol Biol Cell
PubMed ID9436999
Positively charged Nanogold (Nanoprobes, Stony Brook, NY) has been developed as a new marker to follow the endocytic pathway in yeast. Positively charged Nanogold binds extensively to the surface of yeast spheroplasts and is internalized in an energy-dependent manner. Internalization of gold is blocked in the end3 mutant. During a ... More
Efficient immunocytochemical labeling of leukocyte microtubules with FluoroNanogold: an important tool for correlative microscopy.
We tested the immunoprobe FluoroNanogold (FNG) for its utility as an immunocytochemical labeling reagent. This immunoprobe consists of a 1.4-nm gold particle to which a specific Fab' fragment and a fluorochrome are conjugated. We employed the microtubules (MTs) of human phagocytic leukocytes as a model system for testing the usefulness ... More
Electron microscopic visualization of insulin translocation into the cytoplasm and nuclei of intact H35 hepatoma cells using covalently linked Nanogold-insulin.
AuthorsShah N, Zhang S, Harada S, Smith RM, Jarett L
JournalEndocrinology
PubMed ID7789307
Insulin affects numerous metabolic processes as well as nuclear events such as gene transcription. Our previous ultrastructural and biochemical studies demonstrated insulin accumulation in nuclei of cultured and rapidly proliferating cells, and biochemical evidence suggested that insulin entered the cell cytoplasm before accumulating in the nucleus. The present study was ... More
Non-radioactive localization of substance P binding sites in rat brain and spinal cord using peptides labeled with 1.4-nm gold particles.
AuthorsSegond von Banchet G, Heppelmann B
JournalJ Histochem Cytochem
PubMed ID7542681
We present here a new method for non-radioactive labeling of substance P (SP) to demonstrate the distribution of its binding sites in histological sections. The peptide was labeled at the primary amino group with a 1.4-nm gold particle. In Western blots of membrane fractions of rat spinal cord, specific binding ... More
Piezoelectric quartz crystal impedance and electrochemical impedance study of HSA-diazepam interaction by nanogold-structured sensor.
AuthorsLong Y, Nie L, Chen J, Yao S
JournalJ Colloid Interface Sci
PubMed ID12804891
Human serum albumin (HSA) was immobilized on the surface of colloidal Au and exposed to diazepam. Colloidal Au were at first self-assembled on the gold electrode through the thiol groups of a 1,6-hexanedithiol monolayer. The real-time course of the resonant frequency and equivalent circuit parameters of the sensor during the ... More
Simultaneous detection of RNA and protein by in situ hybridization and immunological staining.
AuthorsNagaso H, Murata T, Day N, Yokoyama KK
JournalJ Histochem Cytochem
PubMed ID11511686
Proteinase K is widely used in methods for detection of transcripts in biological specimens by in situ hybridization (ISH). However, treatment with proteinase K hampers detection of RNA and protein simultaneously. We have developed a method for double staining of transcripts and proteins by ISH and IHC staining in imaginal ... More
Real-time investigation of the interaction between primaquine phosphate and bovine serum albumin (BSA) by piezoelectric quartz crystal impedance analysis.
AuthorsLong Y, Chen J, Zhang Z, Yao S
JournalJ Biotechnol
PubMed ID14511914
Real-time investigation of the interaction between primaquine phosphate and bovine serum albumin by the piezoelectric quartz crystal impedance (PQCI) analysis was carried out for the first time. Three kinds of electrodes were investigated. Compared with bare gold (Au) electrode, the gold electrode self-assembled of nanogold colloids exhibits maintained biocompatibility, increased ... More
The enhancement effect of gold nanoparticles as a surface modifier on DNA sensor sensitivity.
AuthorsLiu T, Tang J, Jiang L
JournalBiochem Biophys Res Commun
PubMed ID14672689
The enhancement of a single strain DNA probe linked to the sensor surface is of crucial importance in DNA molecule recognition. By means of nanogold modification of the sensor surface in addition to the nanogold amplifier, DNA detection sensitivity higher than 10(-16)mol/L was obtained in a Quartz Crystal microbalance (QCM) ... More
Using laser scanning confocal microscopy as a guide for electron microscopic study: a simple method for correlation of light and electron microscopy.
AuthorsSun XJ, Tolbert LP, Hildebrand JG
JournalJ Histochem Cytochem
PubMed ID7868862
'Anatomic study of synaptic connections in the nervous system is laborious and difficult, especially when neurons are large or have fine branches embedded among many other processes. Although electron microscopy provides a powerful tool for such study, the correlation of light microscopic appearance and electron microscopic detail is very time-consuming. ... More
Immunogold localization of inositol 1,4,5-trisphosphate receptors and characterization of ultrastructural features of the sarcoplasmic reticulum in phasic and tonic smooth muscle.
AuthorsNixon GF, Mignery GA, Somlyo AV
JournalJ Muscle Res Cell Motil
PubMed ID7706424
'Although agonist stimulation leads to an increase in inositol 1,4,5-trisphosphate (InsP3) and decreased calcium in peripherally and centrally located sarcoplasmic reticulum in smooth muscle, the distribution of InsP3 receptors is unknown. InsP3 receptor and the calcium binding protein, calsequestrin were localized by immunolabelling in a tonic and a phasic smooth ... More
Localization of core spindle pole body (SPB) components during SPB duplication in Saccharomyces cerevisiae.
AuthorsAdams IR, Kilmartin JV
JournalJ Cell Biol
PubMed ID10330408
'We have examined the process of spindle pole body (SPB) duplication in Saccharomyces cerevisiae by electron microscopy and found several stages. These include the assembly, probably from the satellite, of a large plaque-like structure, the duplication plaque, on the cytoplasmic face of the half-bridge and its insertion into the nuclear ... More
Identification by differential display of a hypertonicity-inducible inward rectifier potassium channel highly expressed in chloride cells.
AuthorsSuzuki Y, Itakura M, Kashiwagi M, Nakamura N, Matsuki T, Sakuta H, Naito N, Takano K, Fujita T, Hirose S
JournalJ Biol Chem
PubMed ID10196230
'By using differential mRNA display to monitor the molecular alterations associated with adaptation of euryhaline eels to different salinities, we identified a cDNA fragment strongly induced in seawater eel gills. Cloning of a full-length cDNA and its expression in COS-7 cells indicated that the clone codes for an inward rectifier ... More
Subcellular localization of SV2 and other secretory vesicle components in PC12 cells by an efficient method of preembedding EM immunocytochemistry for cell cultures.
AuthorsTanner VA, Ploug T, Tao-Cheng JH
JournalJ Histochem Cytochem
PubMed ID8985140
'We demonstrated the subcellular localization of SV2, a transmembrane protein associated with neuroendocrine secretory vesicles, in NGF-treated PC12 cells by preembedding EM immunocytochemistry (ICC), using a small gold probe followed by silver enhancement. The use of a multiwell chamber slide substantially improved the efficiency of the preembedding EM ICC procedures ... More
Immunolocalization of a Drosophila phosphatidylinositol transfer protein (rdgB) in normal and rdgA mutant photoreceptor cells with special reference to the subrhabdomeric cisternae.
AuthorsSuzuki E, Hirosawa K
JournalJ Electron Microsc (Tokyo)
PubMed ID7996077
'Distribution of rdgB protein, which was recently shown to be a Drosophila phosphatidylinositol transfer protein, was studied in the photoreceptor cells of compound eyes of normal and rdgAPC47 mutant of Drosophila melanogaster by immunoelectron microscopy using (1) pre-embedding HRP staining, (2) pre-embedding NANOGOLD labeling followed by siliver enhancement, (3) and ... More
New frontiers in gold labeling.
AuthorsHainfeld JF, Powell RD
JournalJ Histochem Cytochem
PubMed ID10727288
'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
Subsynaptic segregation of metabotropic and ionotropic glutamate receptors as revealed by immunogold localization.
AuthorsNusser Z, Mulvihill E, Streit P, Somogyi P
JournalNeuroscience
PubMed ID7969918
'Glutamate is a major neurotransmitter in the brain that acts both through fast ionotropic receptors and through slower metabotropic receptors coupled to G proteins. Both receptors are present throughout the somatodendritic domain of neurons as shown by immunohistochemical and patch clamp recording studies. Immunogold labelling revealed a concentration of metabotropic ... More
KIFC3, a microtubule minus end-directed motor for the apical transport of annexin XIIIb-associated Triton-insoluble membranes.
AuthorsNoda Y, Okada Y, Saito N, Setou M, Xu Y, Zhang Z, Hirokawa N
JournalJ Cell Biol
PubMed ID11581287
'We have identified and characterized a COOH-terminal motor domain-type kinesin superfamily protein (KIFC), KIFC3, in the kidney. KIFC3 is a minus end-directed microtubule motor protein, therefore it accumulates in regions where minus ends of microtubules assemble. In polarized epithelial cells, KIFC3 is localized on membrane organelles immediately beneath the apical ... More
A practical technique to postfix nanogold-immunolabeled specimens with osmium and to embed them in Epon for electron microscopy.
AuthorsSawada H, Esaki M
JournalJ Histochem Cytochem
PubMed ID10727291
'Nanogold is a tiny gold probe, freely diffusible in cells and tissues, and is suitable for pre-embedding immunohistochemistry. However, it is necessary to develop Nanogold to a larger size so that it can be observed by conventional transmission electron microscopy. Silver enhancement is usually used for visualizing Nanogold, but the ... More
ERG30, a VAP-33-related protein, functions in protein transport mediated by COPI vesicles.
AuthorsSoussan L, Burakov D, Daniels MP, Toister-Achituv M, Porat A, Yarden Y, Elazar Z
JournalJ Cell Biol
PubMed ID10427086
'Intracellular transport of newly synthesized and mature proteins via vesicles is controlled by a large group of proteins. Here we describe a ubiquitous rat protein-endoplasmic reticulum (ER) and Golgi 30-kD protein (ERG30)-which shares structural characteristics with VAP-33, a 33-kD protein from Aplysia californica which was shown to interact with the ... More
Expression of guanylin in "pars tuberalis-specific cells" and gonadotrophs of rat adenohypophysis.
AuthorsD'Este L, Kulaksiz H, Rausch U, Vaccaro R, Wenger T, Tokunaga Y, Renda TG, Cetin Y
JournalProc Natl Acad Sci U S A
PubMed ID10655496
'The intestinal peptide guanylin regulates the electrolyte/water transport in the gastrointestinal epithelium by paracrine/luminocrine mechanisms. Because guanylin also circulates in the blood, we investigated the rat hypothalamo-pituitary region for expression and cellular localization of this peptide. Reverse transcriptase-PCR analyses with guanylin-specific primers revealed expression of the peptide in the pars ... More
Immunogold labeling of chromosomes for scanning electron microscopy: a closer look at phosphorylated histone H3 in mitotic metaphase chromosomes of Hordeum vulgare.
AuthorsSchroeder-Reiter E, Houben A, Wanner G
JournalChromosome Res
PubMed ID14516067
'High-resolution detection of phosphorylated histone H3 at serine 10 in mitotic barley chromosomes for scanning electron microscopy was shown using a novel application of indirect immunogold labeling with Nanogold. This method permits localization and quantification of signals in a three-dimensional context. Because the chromosome structure is well preserved, characterization of ... More
A polypeptide bound by the chaperonin groEL is localized within a central cavity.
AuthorsBraig K, Simon M, Furuya F, Hainfeld JF, Horwich AL
JournalProc Natl Acad Sci U S A
PubMed ID8097882
'Chaperonins are oligomeric protein complexes that play an essential role in the cell, mediating ATP-dependent polypeptide chain folding in a variety of cellular compartments. They appear to bind early folding intermediates, preventing their aggregation; in the presence of MgATP and a cochaperonin, bound polypeptides are released in a stepwise manner, ... More
Ultrastructural localization of the vascular permeability factor/vascular endothelial growth factor (VPF/VEGF) receptor-2 (FLK-1, KDR) in normal mouse kidney and in the hyperpermeable vessels induced by VPF/VEGF-expressing tumors and adenoviral vectors.
AuthorsFeng D, Nagy JA, Brekken RA, Pettersson A, Manseau EJ, Pyne K, Mulligan R, Thorpe PE, Dvorak HF, Dvorak AM
JournalJ Histochem Cytochem
PubMed ID10727296
'Vascular permeability factor/vascular endothelial growth factor (VPF/VEGF) interacts with two high-affinity tyrosine kinase receptors, VEGFR-1 and VEGFR-2, to increase microvascular permeability and induce angiogenesis. Both receptors are selectively expressed by vascular endothelial cells and are strikingly increased in tumor vessels. We used a specific antibody to localize VEGFR-2 (FLK-1, KDR) ... More
Group I metabotropic glutamate receptors at GABAergic synapses in monkeys.
AuthorsHanson JE, Smith Y
JournalJ Neurosci
PubMed ID10414977
'Recent data showed that group I metabotropic glutamate receptors (mGluRs) are located perisynaptic to the postsynaptic specializations of asymmetric glutamatergic synapses in the cerebellum and hippocampus in rats. In the present study, we used immunogold labeling to elucidate the subsynaptic localization of group I mGluRs (mGluR1a and mGluR5) in the ... More
Vitronectin in the cytoplasm of Leydig cells in the rat testis.
AuthorsSawada H, Sugawara I, Kitami A, Hayashi M
JournalBiol Reprod
PubMed ID8837997
'Vitronectin is a multifunctional protein present in serum and in extracellular matrices, but its localization has not been fully elucidated. In the present study, immunoblotting with anti-rat vitronectin antibody showed that rat testes contained vitronectin in both Triton-soluble and -insoluble fractions. Immunofluorescence microscopy demonstrated that this immunoreactivity was localized mainly ... More
Nuclear and neuropil aggregates in Huntington's disease: relationship to neuropathology.
AuthorsGutekunst CA, Li SH, Yi H, Mulroy JS, Kuemmerle S, Jones R, Rye D, Ferrante RJ, Hersch SM, Li XJ
JournalJ Neurosci
PubMed ID10087066
'The data we report in this study concern the types, location, numbers, forms, and composition of microscopic huntingtin aggregates in brain tissues from humans with different grades of Huntington''s disease (HD). We have developed a fusion protein antibody against the first 256 amino acids that preferentially recognizes aggregated huntingtin and ... More
Salient features of synaptic organisation in the cerebral cortex.
AuthorsSomogyi P, Tamás G, Lujan R, Buhl EH
JournalBrain Res Brain Res Rev
PubMed ID9651498
'The neuronal and synaptic organisation of the cerebral cortex appears exceedingly complex, and the definition of a basic cortical circuit in terms of defined classes of cells and connections is necessary to facilitate progress of its analysis. During the last two decades quantitative studies of the synaptic connectivity of identified ... More
Expression patterns of transmembrane and released forms of neuregulin during spinal cord and neuromuscular synapse development.
'We mapped the distribution of neuregulin and its transmembrane precursor in developing, embryonic chick and mouse spinal cord. Neuregulin mRNA and protein were expressed in motor and sensory neurons shortly after their birth and levels steadily increased during development. Expression of the neuregulin precursor was highest in motor and sensory ... More
Differential synaptic localization of two major gamma-aminobutyric acid type A receptor alpha subunits on hippocampal pyramidal cells.
AuthorsNusser Z, Sieghart W, Benke D, Fritschy JM, Somogyi P
JournalProc Natl Acad Sci U S A
PubMed ID8876241
'Hippocampal pyramidal cells, receiving domain specific GABAergic inputs, express up to 10 different subunits of the gamma-aminobutyric acid type A (GABAA) receptor, but only 3 different subunits are needed to form a functional pentameric channel. We have tested the hypothesis that some subunits are selectively located at subsets of GABAergic ... More
Target-cell-specific concentration of a metabotropic glutamate receptor in the presynaptic active zone.
AuthorsShigemoto R, Kulik A, Roberts JD, Ohishi H, Nusser Z, Kaneko T, Somogyi P
JournalNature
PubMed ID8632825
'The probability of synaptic neurotransmitter release from nerve terminals is regulated by presynaptic receptors responding to transmitters released from the same nerve terminal or from terminals of other neurons. The release of glutamate, the major excitatory neurotransmitter, is suppressed by presynaptic autoreceptors. Here we show that a metabotropic glutamate receptor ... More
Bax and Bak coalesce into novel mitochondria-associated clusters during apoptosis.
AuthorsNechushtan A, Smith CL, Lamensdorf I, Yoon SH, Youle RJ
JournalJ Cell Biol
PubMed ID11402069
'Bax is a member of the Bcl-2 family of proteins known to regulate mitochondria-dependent programmed cell death. Early in apoptosis, Bax translocates from the cytosol to the mitochondrial membrane. We have identified by confocal and electron microscopy a novel step in the Bax proapoptotic mechanism immediately subsequent to mitochondrial translocation. ... More
Organization of AMPA receptor subunits at a glutamate synapse: a quantitative immunogold analysis of hair cell synapses in the rat organ of Corti.
AuthorsMatsubara A, Laake JH, Davanger S, Usami S, Ottersen OP
JournalJ Neurosci
PubMed ID8699256
'Sensitive and high-resolution immunocytochemical procedures were used to investigate the spatial organization of AMPA receptor subunits (GluR1-4) at the synapse between the inner hair cells and the afferent dendrites in the rat organ of Corti. This is a synapse with special functional properties and with a presynaptic dense body that ... 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
Repeat motifs of tau bind to the insides of microtubules in the absence of taxol.
AuthorsKar S, Fan J, Smith MJ, Goedert M, Amos LA
JournalEMBO J
PubMed ID12505985
'The tau family of microtubule-associated proteins has a microtubule-binding domain which includes three or four conserved sequence repeats. Pelleting assays show that when tubulin and tau are co- assembled into microtubules, the presence of taxol reduces the amount of tau incorporated. In the absence of taxol, strong binding sites for ... More
Localization of the N-terminal domain of the low density lipoprotein receptor.
AuthorsJeon H, Shipley GG
JournalJ Biol Chem
PubMed ID10889195
'The low density lipoprotein (LDL) receptor is a transmembrane glycoprotein performing "receptor-mediated endocytosis" of cholesterol-rich lipoproteins. At the N terminus, the LDL receptor has modular cysteine-rich repeats in both the ligand binding domain and the epidermal growth factor (EGF) precursor homology domain. Each repeat contains six disulfide-bonded cysteine residues, and ... More
Metallosomes.
AuthorsHainfeld JF, Furuya FR, Powell RD
JournalJ Struct Biol
PubMed ID10527904
'Structures and ordered arrays containing organometallic particles have potential application in nanofabrication, smaller computer components, optical devices, sensors, and membrane probes and as detection agents. Here, we describe construction of gold clusters covalently attached to lipids and their use in forming typical lipid structures: micelles, liposomes ("metallosomes"), and sheets on ... More
Vesicle-reconstituted low density lipoprotein receptor. Visualization by cryoelectron microscopy.
AuthorsJeon H, Shipley GG
JournalJ Biol Chem
PubMed ID10889196
'The low density lipoprotein (LDL) receptor is a key protein for maintaining cellular cholesterol homeostasis by binding cholesterol-rich lipoproteins through their apoB and apoE apoproteins. The LDL receptor is a transmembrane glycoprotein of M(r) approximately 115 kDa; based on its primary sequence, five distinct structural domains have been identified (Yamamoto, ... 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
Highly sensitive DNA, RNA and antigen detection methods: streptavidin-Nanogold-silver staining.
AuthorsHauser-Kronberger C
JournalCell Vis
PubMed ID9660736
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
Structural studies of the detergent-solubilized and vesicle-reconstituted insulin receptor.
AuthorsWoldin CN, Hing FS, Lee J, Pilch PF, Shipley GG
JournalJ Biol Chem
PubMed ID10574975
'Insulin binding to the insulin receptor initiates a cascade of cellular events that are responsible for regulating cell metabolism, proliferation, and growth. We have investigated the structure of the purified, functionally active, human insulin receptor using negative stain and cryo-electron microscopy. Visualization of the detergent-solubilized and vesicle-reconstituted receptor shows the ... 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
Organization of 'nanocrystal molecules' using DNA.
'Patterning matter on the nanometre scale is an important objective of current materials chemistry and physics. It is driven by both the need to further miniaturize electronic components and the fact that at the nanometre scale, materials properties are strongly size-dependent and thus can be tuned sensitively. In nanoscale crystals, ... More
Three-dimensional reconstruction of a complex of human alpha 2-macroglobulin with monomaleimido Nanogold (Au1.4nm) embedded in ice.
AuthorsBoisset N, Grassucci R, Penczek P, Delain E, Pochon F, Frank J, Lamy JN
JournalJ Struct Biol
PubMed ID1283687
'Cysteine 949 and glutamine 952 are known to be part of the thiol ester site of each of the four subunits of human alpha 2-macroglobulin (alpha 2M). The hydrolysis of this thiol ester bound to methylamine results in the incorporation of the amine and liberation of a free sulfhydryl group ... More
Labelling and characterisation of gamma-aminobutyric acidA receptor subunit-specific antibodies with monomaleimido Nanogold.
AuthorsRayner SL, Stephenson FA
JournalBiochem Soc Trans
PubMed ID9388760
Three-dimensional reconstruction of human alpha 2-macroglobulin and refinement of the localization of thiol ester bonds with monomaleimido nanogold.
AuthorsBoisset N, Penczek P, Pochon F, Frank J, Lamy J
JournalAnn N Y Acad Sci
PubMed ID7524399
The efficacy of in situ PCR, CARD and nanogold systems for gene detection.
AuthorsNuovo GJ
JournalAm J Pathol
PubMed ID9327752
Ultra-small gold particles and silver enhancement as a detection system in immunolabeling and in situ hybridization experiments.
AuthorsHumbel BM, Sibon OC, Stierhof YD, Schwarz H
JournalJ Histochem Cytochem
PubMed ID7608530
Confocal epipolarization microscopy of gold probes in plant cells and protoplasts.
AuthorsGriffing LR, Villanueva MA, Taylor J, Moon S
JournalMethods Cell Biol
PubMed ID8531749
Silver-enhanced immunogold labeling of calcium-ATPase in sarcoplasmic reticulum of skeletal muscle.
AuthorsKrenács T, Dux L
JournalJ Histochem Cytochem
PubMed ID8014481
Immunogold labeling in scanning electron microscopy.
AuthorsHermann R, Walther P, Müller M
JournalHistochem Cell Biol
PubMed ID8858365
Enhanced labeling efficiency using ultrasmall immunogold probes: immunocytochemistry.
AuthorsRobinson JM, Takizawa T, Vandré DD
JournalJ Histochem Cytochem
PubMed ID10727290
Detection of antigen-antibody interactions in immunocytochemistry relies on a reporter system. The most commonly employed reporter systems used are fluorochromes, enzymes, and particulate probes. This article considers the advantages and disadvantages associated with ultrasmall immunogold particles as the reporter system in immunocytochemical applications. ... More
Single-mismatch detection using gold-quenched fluorescent oligonucleotides.
AuthorsDubertret B, Calame M, Libchaber AJ
JournalNat Biotechnol
PubMed ID11283596
Here we describe a hybrid material composed of a single-stranded DNA (ssDNA) molecule, a 1.4 nm diameter gold nanoparticle, and a fluorophore that is highly quenched by the nanoparticle through a distance-dependent process. The fluorescence of this hybrid molecule increases by a factor of as much as several thousand as ... 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
Use of nanogold followed by silver enhancement and gold toning for preembedding immunolocalization in osmium-fixed, Epon-embedded tissues.
AuthorsSawada H, Esaki M
JournalJ Electron Microsc (Tokyo)
PubMed ID7722428
A reliable, sensitive, high-resolution method with good structural visualization for preembedding immunoelectron microscopy was proposed. The present technique involves the following processes. Immunolabeling of cryostat sections with primary antibodies and with nanogolds, silver intensification for visualization of nanogold secondary antibodies, gold-toning for stabilization of silver shells, and osmium postfixation and ... More
Conformational constraints in protein degradation by the 20S proteasome.
AuthorsWenzel T, Baumeister W
JournalNat Struct Biol
PubMed ID7773788
Conformationally stabilized peptides and unfolding intermediates of bovine alpha-lactalbumin have been used to define the degree of unfolding required for degradation by 20S proteasomes. It appears that complete unfolding and the absence of disulphide bonds are prerequisites for degradation, suggesting that a relatively narrow opening controls access to the inner ... More
CD95 signaling via ceramide-rich membrane rafts.
AuthorsGrassme H, Jekle A, Riehle A, Schwarz H, Berger J, Sandhoff K, Kolesnick R, Gulbins E
JournalJ Biol Chem
PubMed ID11279185
Clustering seems to be employed by many receptors for transmembrane signaling. Here, we show that acid sphingomyelinase (ASM)-released ceramide is essential for clustering of CD95. In vitro and in vivo, extracellularly orientated ceramide, released upon CD95-triggered translocation of ASM to the plasma membrane outer surface, enabled clustering of CD95 in ... More
Localization of Rabphilin-3A on the synaptic vesicle.
AuthorsMizoguchi A, Yano Y, Hamaguchi H, Yanagida H, Ide C, Zahraoui A, Shirataki H, Sasaki T, Takai Y
JournalBiochem Biophys Res Commun
PubMed ID8060298
Rabphilin-3A is a putative target protein for Rab3A small GTP-binding protein which is implicated in neurotransmitter release. Rabphilin-3A is expressed mainly in brain, but its subcellular localization remains to be clarified. Immunohistochemical analysis has revealed that Rabphilin-3A is most abundant in the synaptic area of the rat cerebellum, retina, and ... More
Correlative microscopy using FluoroNanogold on ultrathin cryosections. Proof of principle.
AuthorsTakizawa T, Suzuki K, Robinson JM
JournalJ Histochem Cytochem
PubMed ID9742065
We demonstrate a fluorescent ultrasmall immunogold probe, FluoroNanogold (FNG), to be a versatile reporter system for immunocytochemical labeling of ultrathin cryosections. FNG-labeled molecules in the same ultrathin cryosections can be resolved by two imaging techniques (i.e., fluorescence and electron microscopy). Lactoferrin, a marker protein for the specific granules in human ... More
Electron microscopy of alpha 2-macroglobulin with a thiol ester bound ligand.
AuthorsBoisset N, Pochon F, Chwetzoff S, Barray M, Delain E, Lamy J
JournalJ Struct Biol
PubMed ID1282356
In order to covalently bind the hydrolyzed thiol ester groups of the human alpha 2-macroglobulin (alpha 2M) transformed by methylamine, the phospholipase A2 (PLA2), a small enzyme (M(r) = 13,000) from Naja nigricollis snake venom was activated by succinimidyl 4-(maleimidomethyl)cyclohexane-1-carboxylate (SMCC). Average images determined from electron micrographs of the methylamine-transformed ... More
Monomaleimidogold labeling of the gamma subunit of the Escherichia coli F1 ATPase examined by cryoelectron microscopy.
AuthorsWilkens S, Capaldi RA
JournalArch Biochem Biophys
PubMed ID1444442
A novel approach for locating sites of interest in a protein complex has been developed using monomaleimidonanogold (MMN). The Escherichia coli F1 ATPase, when prepared without the delta subunit, contains only a single reactive cysteine on one of the three copies of the alpha subunit. This site was reacted with ... More
A 1.4-nm gold cluster covalently attached to antibodies improves immunolabeling.
AuthorsHainfeld JF, Furuya FR
JournalJ Histochem Cytochem
PubMed ID1552162
A large gold cluster (Au1.4nm) was covalently coupled to IgG and Fab' fragments. Its gold core is 1.4 nm in diameter and the Fab'-Au1.4nm immunoconjugate is the smallest gold immunoprobe that can be seen directly in the conventional electron microscope. It is useful in high-resolution immunolabeling, providing a resolution of ... More
Subcellular localization of estradiol receptor in MCF7 cells studied with nanogold-labelled antibody fragments.
AuthorsKessels MM, Qualmann B, Thole HH, Sierralta WD
JournalEur J Histochem
PubMed ID10068898
Ultrastructural localization studies of estradiol receptor in hormone-deprived and hormone-stimulated MCF7 cells were done using F(ab') fragments of three different antibodies (#402, 13H2, HT277) covalently linked to nanogold. These ultra-small, non-charged immunoreagents, combined with a size-enlargement by silver enhancement, localized estradiol receptor in both nuclear and cytoplasmic areas of non-stimulated ... More
Ni-NTA-gold clusters target His-tagged proteins.
AuthorsHainfeld JF, Liu W, Halsey CM, Freimuth P, Powell RD
JournalJ Struct Biol
PubMed ID10527908
Addition of six histidines to recombinant proteins has proved useful in their purification by nickel-affinity columns. This technology was adapted by synthesizing the chelator for nickel (nitrilotriacetic acid, NTA) onto the surface of gold clusters. These Ni-NTA-gold clusters were shown to specifically target the 6His region of tagged proteins. Results ... More
The metabotropic glutamate receptor (mGluR1 alpha) is concentrated at perisynaptic membrane of neuronal subpopulations as detected by immunogold reaction.
AuthorsBaude A, Nusser Z, Roberts JD, Mulvihill E, McIlhinney RA, Somogyi P
JournalNeuron
PubMed ID8104433
An antiserum to mGluR1 alpha labeled a 160 kd protein in immunoblots of membranes derived from rat brain or cells transfected with mGluR1 alpha. Immunoreactivity for mGluR1 alpha was present in discrete subpopulations of neurons. The GABAergic neurons of the cerebellar cortex were strongly immunoreactive; only some Golgi cells were ... 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
Cryoelectron microscopy of low density lipoprotein in vitreous ice.
AuthorsSpin JM, Atkinson D
JournalBiophys J
PubMed ID7612855
In this report, images of low density lipoprotein (LDL) in vitreous ice at approximately 30 A resolution are presented. These images show that LDL is a quasi-spherical particle, approximately 220-240 A in diameter, with a region of low density (lipid) surrounded by a ring (in projection) of high density believed ... More
Colorimetric detection of protein microarrays based on nanogold probe coupled with silver enhancement.
AuthorsLiang RQ, Tan CY, Ruan KC
JournalJ Immunol Methods
PubMed ID14980430
This work presents a method for analyzing protein microarrays using a colorimetric nanogold probe coupled with silver enhancement (gold-silver detection). In this method, the gold nanoparticles were introduced to the microarray by the specific binding of the gold-conjugated antibodies or streptavidins and then coupled with silver enhancement to produce black ... More
A covalent fluorescent-gold immunoprobe: simultaneous detection of a pre-mRNA splicing factor by light and electron microscopy.
Immunoprobes that combine a fluorescent label with a 1.4-nm gold cluster compound have been prepared by covalent conjugation with polyclonal antibody Fab' fragments. These new immunoconjugates allow the collection of two complementary sets of data, from fluorescence and electron microscopy, from a single labeling experiment. We find that incorporation of ... More
Peculiar distribution of fodrin in fat-storing cells.
AuthorsAoki T, Hagiwara H, Fujimoto T
JournalExp Cell Res
PubMed ID9260899
Fat-storing cells (FSCs) show unique morphology containing many lipid droplets in the cytoplasm. In this study, we found that a membrane skeletal protein, fodrin, shows peculiar distribution in FSCs of rat liver. By immunofluorescence microscopy of FSCs in culture, intense labeling for fodrin was seen as coarse filaments in the ... More
Ultrathin cryosections: an important tool for immunofluorescence and correlative microscopy.
AuthorsTakizawa T, Robinson JM
JournalJ Histochem Cytochem
PubMed ID12754282
Here we show that ultrathin cryosections of placental tissue can be used as a substrate in immunofluorescence experiments. A high degree of spatial resolution can be achieved in these preparations because there is essentially no out-of-focus fluorescence. Therefore, immunofluorescence microscopy using ultrathin cryosections provides a very useful method for determining ... More
Ultrastructural localization of gustducin immunoreactivity in microvilli of type II taste cells in the rat.
Gustducin is a transducin-like G protein (guanine nucleotide-binding protein) that is expressed in taste bud cells. Gustducin is believed to be involved in bitter and possibly sweet taste transduction. In the present study, we demonstrate that a subset of type II cells displays immunoreactivity to antisera directed against gustducin in ... More
High performance Nanogold-silver in situ hybridisation.
AuthorsHacker GW
JournalEur J Histochem
PubMed ID9728288
Conventional in situ hybridisation (ISH) usually requires the presence of at least 10-50 copies of the nucleic acid sequence in question per cell. In situ PCR has been proposed as an alternative method, which may yield single-copy sensitivity, but shows a relatively high rate of false-negative or even false-positive reactions. ... More
Signal enhancement at the electron microscopic level using Nanogold and gold-based autometallography.
AuthorsWeipoltshammer K, Schöfer C, Almeder M, Wachtler F
JournalHistochem Cell Biol
PubMed ID11201611
Immunoelectron microscopy using ultrasmall gold markers is a very sensitive method to detect molecules at high resolution. In order to discriminate the gold particles in the electron microscope, enlargement of gold particles is necessary. So far, mostly silver ions were used for deposition onto the surface of gold grains. In ... More
Localization of a metabotropic glutamate receptor, mGluR7, in axon terminals of presumed nociceptive, primary afferent fibers in the superficial layers of the spinal dorsal horn: an electron microscope study in the rat.
AuthorsLi H, Ohishi H, Kinoshita A, Shigemoto R, Nomura S, Mizuno N
JournalNeurosci Lett
PubMed ID9080455
It was examined electron microscopically in the rat if a metabotropic glutamate receptor, mGluR7, might be localized in axon terminals of nociceptive, primary afferent fibers in laminae I and II of the spinal dorsal horn. Nociceptive nature of axon terminals showing mGluR7-like immunoreactivity (mGluR7-LI) was indicated by binding to the ... More
Cloning and characterization of a novel membrane-associated antigenic protein of Helicobacter pylori.
AuthorsYoshida M, Wakatsuki Y, Kobayashi Y, Itoh T, Murakami K, Mizoguchi A, Usui T, Chiba T, Kita T
JournalInfect Immun
PubMed ID9864228
Infection by Helicobacter pylori, a noninvasive bacterium, induces chronic leukocyte infiltration in the stomach by still largely unknown molecular mechanisms. We investigated the possibility that a membrane protein of H. pylori induces an inflammatory reaction in the subepithelial tissue of the stomach. By generating an expression library of H. pylori ... More
Differences in synaptic GABA(A) receptor number underlie variation in GABA mini amplitude.
AuthorsNusser Z, Cull-Candy S, Farrant M
JournalNeuron
PubMed ID9331359
In many neurons, responses to individual quanta of transmitter exhibit large variations in amplitude. The origin of this variability, although central to our understanding of synaptic transmission and plasticity, remains controversial. To examine the relationship between quantal amplitude and postsynaptic receptor number, we adopted a novel approach, combining patch-clamp recording ... More
Structure of the Escherichia coli ATP synthase and role of the gamma and epsilon subunits in coupling catalytic site and proton channeling functions.
AuthorsCapaldi RA, Aggeler R, Gogol EP, Wilkens S
JournalJ Bioenerg Biomembr
PubMed ID1429536
The structure of the Escherichia coli ATP synthase has been studied by electron microscopy and a model developed in which the alpha and beta subunits of the F1 part are arranged hexagonally (in top view) alternating with one another and surrounding a central cavity of around 35 A at its ... More
The use of electron microscopic immunocytochemistry with silver-enhanced 1.4-nm gold particles to localize GAD in the cerebellar nuclei.
AuthorsGilerovitch HG, Bishop GA, King JS, Burry RW
JournalJ Histochem Cytochem
PubMed ID7868863
Silver enhancement of small gold particles can be used with pre-embedding immunocytochemistry to analyze the distribution of label over cell organelles. We have developed a method that improves tissue morphology, has good penetration of reagents, and allows greater control of silver enhancement of 1.4-nm gold. In this study we analyzed ... More
Bidirectional amyloid fiber growth for a yeast prion determinant.
The polymerization of many amyloids is a two-stage process initiated by the formation of a seeding nucleus or protofibril. Soluble protein then assembles with these nuclei to form amyloid fibers. Whether fiber growth is bidirectional or unidirectional has been determined for two amyloids. In these cases, bidirectional growth was established ... More
Use of 1.4-nm immunogold particles for immunocytochemistry on ultra-thin cryosections.
AuthorsTakizawa T, Robinson JM
JournalJ Histochem Cytochem
PubMed ID7983362
We present a new application for the use of small immunogold particles (approximately 1.4-nm diameter) for ultrastructural immunocytochemistry. These small gold particles have been used on ultra-thin cryosections in conjunction with a silver enhancement procedure that does not degrade ultrastructural detail. We have used the human neutrophil as a model ... More
Direct comparison of the sensitivity of enzyme histochemical and immunohistochemical methods: cathepsin B expression in human colorectal mucosa.
AuthorsBleeker FE, Hazen LG, Köhler A, Van Noorden CJ
JournalActa Histochem
PubMed ID10990063
Immunohistochemical localization of the proteinase cathepsin B has been compared directly with localization of cathepsin B activity with a catalytic (enzyme) histochemical method. The 2 approaches demonstrate principally different aspects of an enzyme. The immunohistochemical method localizes the enzyme protein whether it is active or not whereas the catalytic method ... 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
Preparation, use, and enlargement of ultrasmall gold particles in immunoelectron microscopy.
AuthorsBaschong W, Stierhof YD
JournalMicrosc Res Tech
PubMed ID9712164
The introduction of ultrasmall (approximately 1-3 nm) colloidal gold markers in immunoelectron microscopy (IEM) in 1989 has considerably improved the sensitivity of this marker system. Ultrasmall gold markers have opened the field of pre-embedding labeling studies to gold markers without the need of harsh permeabilizing steps. They are recommended for ... More
Immunogold silver staining for light microscopy.
AuthorsLackie PM
JournalHistochem Cell Biol
PubMed ID8858363
The immunogold silver staining method (IGSS) is widely used as a sensitive and specific immunohistochemical visualisation technique. IGSS involves the specific deposition of metallic silver at the site of immunogold labelling and provides a means of visualisation at low magnification by light or electron microscopy. Silver developers for IGSS rapidly ... More
Cellular defects and altered gene expression in PC12 cells stably expressing mutant huntingtin.
AuthorsLi SH, Cheng AL, Li H, Li XJ
JournalJ Neurosci
PubMed ID10377328
Expanded polyglutamine tracts cause huntingtin and other proteins to accumulate and aggregate in neuronal nuclei. Whether the intranuclear aggregation or localization of a polyglutamine protein initiates cellular pathology remains controversial. We established stably transfected pheochromocytoma PC12 cells that express the N-terminal fragment of huntingtin containing 20 (20Q) or 150 (150Q) ... More
Introduction of a novel HRP substrate-Nanogold probe for signal amplification in immunocytochemistry.
AuthorsMayer G, Leone RD, Hainfeld JF, Bendayan M
JournalJ Histochem Cytochem
PubMed ID10727287
Amplification of immunological signals with catalyzed reporter deposition (CARD) allows improved detection of scarce tissue antigens in light and electron microscopy. The technique takes advantage of the oxidation ability of horseradish peroxidase (HRP), in the presence of hydrogen peroxide, to yield the accumulation of one of its specific reporter-tagged substrates. ... More
Relative densities of synaptic and extrasynaptic GABAA receptors on cerebellar granule cells as determined by a quantitative immunogold method.
AuthorsNusser Z, Roberts JD, Baude A, Richards JG, Somogyi P
JournalJ Neurosci
PubMed ID7722639
Ion channels gated by the inhibitory neurotransmitter gamma-aminobutyric acid (GABA) are thought to be located in synaptic junctions, but they have also been found throughout the somatodendritic membrane of neurons independent of synapses. To test whether synaptic junctions are enriched in GABAA receptors, and to determine the relative densities of ... More
Sites of rDNA transcription are widely dispersed through the nucleolus in Pisum sativum and can comprise single genes.
AuthorsThompson WF, Beven AF, Wells B, Shaw PJ
JournalPlant J
PubMed ID9351243
Incorporation by RNA polymerases of BrUTP into both plant root tissue and isolated plant nuclei as a method for localization of the sites of transcription has been used. In this paper pea root tissue was used, and under the conditions employed, nearly all the incorporation occurs in the nucleolus, and ... More
Preparation of plasmid DNA in transfection complexes for fluorescence and electron spectroscopic imaging.
AuthorsMalecki M
JournalScanning Microsc Suppl
PubMed ID9601525
The aim of this project was to develop procedures necessary to study mechanisms of receptor mediated gene transfer by means of integrated microscopy. Plasmid DNA was incorporated into a transfection complex consisting of poly(L)lysine and transferrin to which the nuclear localization signal was conjugated. This complex was presented to cultured ... More
Ultrastructural localization of beta-actin and amphoterin mRNA in cultured cells: application of tyramide signal amplification and comparison of detection methods.
AuthorsPunnonen EL, Fages C, Wartiovaara J, Rauvala H
JournalJ Histochem Cytochem
PubMed ID9857217
We describe a nonradioactive preembedding in situ hybridization protocol using digoxigenin-labeled RNA probes and tyramide signal amplification to increase the sensitivity of detection. The protocol is sensitive enough for electron microscopic localization of endogenous messenger RNAs encoding beta-actin and amphoterin. Three visualization methods were compared: diaminobenzidine enhanced by nickel, Nanogold ... More
The alpha 6 subunit of the GABAA receptor is concentrated in both inhibitory and excitatory synapses on cerebellar granule cells.
AuthorsNusser Z, Sieghart W, Stephenson FA, Somogyi P
JournalJ Neurosci
PubMed ID8613776
Although three distinct subunits seem to be sufficient to form a functional pentameric GABAA receptor channel, cerebellar granule cells express nRNA for nine subunits. They receive GABAergic input from a relatively homogenous population of Golgi cells. It is not known whether all subunits are distributed similarly on the surface of ... More
Colorimetric silver detection of methylation using DNA microarray coupled with linker-PCR.
AuthorsJi M, Hou P, Li S, He N, Lu Z
JournalClin Chim Acta
PubMed ID15026275
BACKGROUND: Aberrant DNA methylation of CpG site is among the earliest and most frequent alterations in cancer. Detection of promoter hypermethylation of cancer-related genes may be useful for cancer diagnosis or the detection of recurrence. Recently, several DNA microarray methods have been developed to detect the methylation status of the ... More
Immunoelectron microscopic localization of phosphoproteins associated with the mitotic spindle.
AuthorsVandré DD, Burry RW
JournalJ Histochem Cytochem
PubMed ID1453002
We examined the immunogold staining of microtubules and microtubule organizing centers using an improved silver-enhancement reagent for small (1-1.4 nm) gold-conjugated secondary antibodies. First, the staining properties of different commercial preparations of gold-labeled antibodies were compared for sample penetration, label uniformity, and labeling density, and Nanogold 1.4-nm gold-conjugated F(ab') was ... More
Binding of amyloid beta protein to the 20 S proteasome.
AuthorsGregori L, Hainfeld JF, Simon MN, Goldgaber D
JournalJ Biol Chem
PubMed ID8995227
Neurodegenerative disorders of aging are characterized by the intraneuronal accumulation of ubiquitin conjugates into tangles and inclusions. Ubiquitin conjugates are degraded by cellular particles known as proteasomes. We have previously shown that amyloid beta protein (Abeta) inhibits proteasomal activity and thereby blocks ubiquitin conjugate degradation. In the present studies, we ... More
Dendritic and postsynaptic protein synthetic machinery.
AuthorsGardiol A, Racca C, Triller A
JournalJ Neurosci
PubMed ID9870948
There is a growing body of evidence that local protein synthesis beneath synapses may provide a novel mechanism underlying plastic phenomena. In vivo and in vitro biochemical data show that dendrites can perform translation and glycosylation. Using antibodies directed against the eukaryotic protein synthetic machinery, we sought to identify the ... 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