Expanding the multicolor capabilities of basic confocal microscopes by employing red and near-infrared quantum dot conjugates.
AuthorsKingeter LM, Schaefer BC,
JournalBMC Biotechnol
PubMed ID19463154
'BACKGROUND: Confocal microscopy is a widely employed methodology in cellular biology, commonly used for investigating biological organization at the cellular and sub-cellular level. Most basic confocal microscopes are equipped to cleanly discriminate no more than four fluorophores in a given sample, limiting the utility of this method for co-localization, co-expression, ... More
Wet electron microscopy with quantum dots.
AuthorsTimp W, Watson N, Sabban A, Zik O, Matsudaira P
JournalBiotechniques
PubMed ID16989089
Wet electron microscopy (EM) is a new imaging method with the potential to allow higher spatial resolution of samples. In contrast to most EM methods, it requires little time to perform and does not require complicated equipment or difficult steps. We used this method on a common murine macrophage cell ... More
Phallotoxin and actin binding assay by fluorescence enhancement.
AuthorsHuang ZJ, Haugland RP, You WM, Haugland RP
JournalAnal Biochem
PubMed ID1595896
The fluorescence of five fluorophores conjugated to phallotoxins was found to be specifically enhanced upon binding to F-actin in a polymerizing buffer. Rhodamine phalloidin had the greatest fluorescence enhancement of ninefold. The fluorescence titration of rhodamine phalloidin by actin was shown to be consistent with stoichiometric binding. The fluorescence enhancement ... More
Activity-dependent tau protein translocation to excitatory synapse is disrupted by exposure to amyloid-Beta oligomers.
AuthorsFrandemiche ML, De Seranno S, Rush T, Borel E, Elie A, Arnal I, Lanté F, Buisson A,
Journal
PubMed ID24760868
Tau is a microtubule-associated protein well known for its stabilization of microtubules in axons. Recently, it has emerged that tau participates in synaptic function as part of the molecular pathway leading to amyloid-beta (Aß)-driven synaptotoxicity in the context of Alzheimer's disease. Here, we report the implication of tau in the ... More
Detection of small differences in actomyosin function using actin labeled with different phalloidin conjugates.
AuthorsBalaz M, Månsson A
JournalAnal Biochem
PubMed ID15745742
This study shows that there is only a negligible difference in actomyosin function in the in vitro motility assay among actin filaments labeled with Rhodamine phalloidin (RhPh), Alexa-488 phalloidin (APh), and biotin-XX phalloidin (BPh). Similar results were obtained at varying ionic strengths (0.02-0.13 M), in the presence of imidazole or ... More
Deformation-enhanced fluctuations in the red cell skeleton with theoretical relations to elasticity, connectivity, and spectrin unfolding.
AuthorsLee JC, Discher DE
JournalBiophys J
PubMed ID11720984
To assess local elasticity in the red cell's spectrin-actin network, nano-particles were tethered to actin nodes and their constrained thermal motions were tracked. Cells were both immobilized and controllably deformed by aspiration into a micropipette. Since the network is well-appreciated as soft, thermal fluctuations even in an unstressed portion of ... More
The ELF -97 phosphatase substrate provides a sensitive, photostable method for labelling cytological targets.
AuthorsParagas VB, Kramer JA, Fox C, Haugland RP, Singer VL
JournalJ Microsc
PubMed ID12000550
We compared fluorescent signals obtained with fluorescein conjugates and the ELF-97 (enzyme-labelled fluorescence) phosphatase substrate [2-(5'-chloro-2-phosphoryloxyphenyl)-6-chloro-4(3H)-quinazolinone] in labelling cytological structures requiring high spatial resolution. Enzymatic cleavage of the ELF-97 phosphatase substrate yields an extremely fine precipitate that remains well localized to the site of enzymatic activity. This precipitate fluoresces bright ... More
Colony-stimulating factor-1 stimulates the formation of multimeric cytosolic complexes of signaling proteins and cytoskeletal components in macrophages.
AuthorsYeung YG, Wang Y, Einstein DB, Lee PS, Stanley ER
JournalJ Biol Chem
PubMed ID9642280
Stimulation of macrophages with colony-stimulating factor-1 (CSF-1) results in the protein tyrosine phosphorylation of the CSF-1 receptor (CSF-1R) and many other, primarily cytosolic, proteins. Stimulation by CSF-1 at 4 degreesC was used to facilitate the purification and identification of the proteins of the cytosolic anti-phosphotyrosine (PY)-reactive fraction (alphaPY-RF) involved in ... More
Caenorhabditis elegans beta-G spectrin is dispensable for establishment of epithelial polarity, but essential for muscular and neuronal function.
AuthorsMoorthy S, Chen L, Bennett V
JournalJ Cell Biol
PubMed ID10811831
The Caenorhabditis elegans genome encodes one alpha spectrin subunit, a beta spectrin subunit (beta-G), and a beta-H spectrin subunit. Our experiments show that the phenotype resulting from the loss of the C. elegans alpha spectrin is reproduced by tandem depletion of both beta-G and beta-H spectrins. We propose that alpha ... More
High resolution mapping of mast cell membranes reveals primary and secondary domains of Fc(epsilon)RI and LAT.
AuthorsWilson BS, Pfeiffer JR, Surviladze Z, Gaudet EA, Oliver JM
JournalJ Cell Biol
PubMed ID11489921
In mast cells, cross-linking the high-affinity IgE receptor (Fc(epsilon)RI) initiates the Lyn-mediated phosphorylation of receptor ITAMs, forming phospho-ITAM binding sites for Syk. Previous immunogold labeling of membrane sheets showed that resting Fc(epsilon)RI colocalize loosely with Lyn, whereas cross-linked Fc(epsilon)RI redistribute into specialized domains (osmiophilic patches) that exclude Lyn, accumulate Syk, ... More
Observing FcepsilonRI signaling from the inside of the mast cell membrane.
AuthorsWilson BS, Pfeiffer JR, Oliver JM
JournalJ Cell Biol
PubMed ID10831616
We have determined the membrane topography of the high-affinity IgE receptor, FcstraightepsilonRI, and its associated tyrosine kinases, Lyn and Syk, by immunogold labeling and transmission electron microscopic (TEM) analysis of membrane sheets prepared from RBL-2H3 mast cells. The method of Sanan and Anderson (Sanan, D.A., and R.G.W. Anderson. 1991. J. ... More
Quantification and localization of phosphorylated myosin I isoforms in Acanthamoeba castellanii.
AuthorsBaines IC, Corigliano-Murphy A, Korn ED
JournalJ Cell Biol
PubMed ID7622560
The actin-activated Mg(2+)-ATPase activities of the three myosin I isoforms in Acanthamoeba castellanii are significantly expressed only after phosphorylation of a single site in the myosin I heavy chain. Synthetic phosphorylated and unphosphorylated peptides corresponding to the phosphorylation site sequences, which differ for the three myosin I isoforms, were used ... More
Deletion of Wiskott-Aldrich syndrome protein triggers Rac2 activity and increased cross-presentation by dendritic cells.
Authors
JournalNat Commun
PubMed ID27425374
Wide-field three-photon excitation in biological samples.
Authors
JournalLight Sci Appl
PubMed ID29152380
The cortical actin network regulates avidity-dependent binding of hyaluronan by the lymphatic vessel endothelial receptor LYVE-1.
Authors
JournalJ Biol Chem
PubMed ID32034091
Thin filament length dysregulation contributes to muscle weakness in nemaline myopathy patients with nebulin deficiency.
Authors
JournalHum Mol Genet
PubMed ID19346529
Nebulin regulates thin filament length, contractility, and Z-disk structure in vivo.
Authors
JournalEMBO J
PubMed ID16902413
Interferon-induced mechanosensing defects impede apoptotic cell clearance in lupus.
Authors
JournalJ Clin Invest
PubMed ID26098211
Biased localization of actin binding proteins by actin filament conformation.
Authors
JournalNat Commun
PubMed ID33239610
Biallelic loss of human CTNNA2, encoding αN-catenin, leads to ARP2/3 complex overactivity and disordered cortical neuronal migration.
Authors
JournalNat Genet
PubMed ID30013181
Acquisition of cellular properties during alveolar formation requires differential activity and distribution of mitochondria.
Authors
JournalElife
PubMed ID35384838
Anillin propels myosin-independent constriction of actin rings.