Parallel detection of antigen-specific T-cell responses by multidimensional encoding of MHC multimers.
AuthorsHadrup SR, Bakker AH, Shu CJ, Andersen RS, van Veluw J, Hombrink P, Castermans E, Thor Straten P, Blank C, Haanen JB, Heemskerk MH, Schumacher TN,
JournalNat Methods
PubMed ID19543285
'The use of fluorescently labeled major histocompatibility complex multimers has become an essential technique for analyzing disease- and therapy-induced T-cell immunity. Whereas classical major histocompatibility complex multimer analyses are well-suited for the detection of immune responses to a few epitopes, limitations on human-subject sample size preclude a comprehensive analysis of ... More
Single-molecule quantum-dot fluorescence resonance energy transfer.
AuthorsHohng S, Ha T
JournalChemphyschem
PubMed ID15884082
'Colloidal semiconductor quantum dots are promising for single-molecule biological imaging due to their outstanding brightness and photostability. As a proof of concept for single-molecule fluorescence resonance energy transfer (FRET) applications, we measured FRET between a single quantum dot and a single organic fluorophore Cy5. DNA Holliday junction dynamics measured with ... More
Analysis of UV-excited fluorochromes by flow cytometry using near-ultraviolet laser diodes.
AuthorsTelford WG
JournalCytometry A
PubMed ID15351984
'INTRODUCTION: Violet laser diodes have become common and reliable laser sources for benchtop flow cytometers. While these lasers are very useful for a variety of violet and some ultraviolet-excited fluorochromes (e.g., DAPI), they do not efficiently excite most UV-stimulated probes. In this study, the next generation of InGaN near-UV laser ... More
The development of quantum dot calibration beads and quantitative multicolor bioassays in flow cytometry and microscopy.
AuthorsWu Y, Campos SK, Lopez GP, Ozbun MA, Sklar LA, Buranda T
JournalAnal Biochem
PubMed ID17397793
'The use of fluorescence calibration beads has been the hallmark of quantitative flow cytometry. It has enabled the direct comparison of interlaboratory data as well as quality control in clinical flow cytometry. In this article, we describe a simple method for producing color-generalizable calibration beads based on streptavidin functionalized quantum ... More
Quantum dot self-assembly for protein detection with sub-picomolar sensitivity.
AuthorsSoman CP, Giorgio TD,
JournalLangmuir
PubMed ID18335969
'A novel approach to sensitive and rapid antigen detection is described. In the presence of a specific antigen, quantum dot-antibody conjugates rapidly self-assemble into agglomerates that are typically more than 1 order of magnitude larger than their individual components. The size distribution of the agglomerated colloids depends on, among other ... More
Separation of bioconjugated quantum dots using capillary electrophoresis.
AuthorsVicente G, Colón LA,
JournalAnal Chem
PubMed ID18278946
'Capillary electrophoresis (CE) with laser-induced fluorescence (LIF) detection was used to separate different bioconjugated CdSe/ZnS quantum dots (QDs). The QD nanocrystals studied were conjugated to the biomolecules streptavidin, biotin, and immunoglobulin G. The bioconjugated QDs showed different electrophoretic mobilities, which appear to depend upon the biomolecule that is attached to ... More
Nanometer distance measurements between multicolor quantum dots.
'Quantum dot dimers made of short double-stranded DNA molecules labeled with different color quantum dots at each end were imaged using multicolor stage-scanning confocal microscopy. This approach eliminates chromatic aberration and color registration issues usually encountered in other multicolor imaging techniques. We demonstrate nanometer accuracy in individual distance measurement by ... More
Quantum dot mediated imaging of atherosclerosis.
AuthorsJayagopal A, Su YR, Blakemore JL, Linton MF, Fazio S, Haselton FR,
JournalNanotechnology
PubMed ID19420562
'The progression of atherosclerosis is associated with leukocyte infiltration within lesions. We describe a technique for the ex vivo imaging of cellular recruitment in atherogenesis which utilizes quantum dots (QD) to color-code different cell types within lesion areas. Spectrally distinct QD were coated with the cell-penetrating peptide maurocalcine to fluorescently-label ... More
Method for multiplex cellular detection of mRNAs using quantum dot fluorescent in situ hybridization.
'The photostability and narrow emission spectra of non-organic quantum dot fluorophores (QDs) make them desirable candidates for fluorescent in situ hybridization (FISH) to study the expression of specific mRNA transcripts. We developed a novel method for direct QD labeling of modified oligonucleotide probes through streptavidin and biotin interactions, as well ... More
Photodynamics of a single quantum dot: fluorescence activation, enhancement, intermittency, and decay.
AuthorsLee SF, Osborne MA
JournalJ Am Chem Soc
PubMed ID17608479
Inflammation in fetal sheep from intra-amniotic injection of Ureaplasma parvum.
AuthorsCollins JJ, Kallapur SG, Knox CL, Nitsos I, Polglase GR, Pillow JJ, Kuypers E, Newnham JP, Jobe AH, Kramer BW,
JournalAm J Physiol Lung Cell Mol Physiol
PubMed ID20935228
Bronchopulmonary dysplasia is associated with chorioamnionitis and fetal lung inflammation. Ureaplasma species are the bacteria most frequently isolated from chorioamnionitis. Very chronic ureaplasma colonization of amniotic fluid causes low-grade lung inflammation and functional lung maturation in fetal sheep. Less is known about shorter exposures of the fetal lung. Therefore, we ... More
Suppression of quantum dot blinking in DTT-doped polymer films.
AuthorsAntelman J, Ebenstein Y, Dertinger T, Michalet X, Weiss S,
JournalJ Phys Chem C Nanomater Interfaces
PubMed ID20161096
In this report we evaluate the emission properties of single quantum dots embedded in a thin, thiol containing polymer film. We report the suppression of quantum dot blinking leading to a continuous photon flux from both organic and water soluble quantum dots and demonstrate their application as robust fluorescent point ... More
Actin restricts FcepsilonRI diffusion and facilitates antigen-induced receptor immobilization.
AuthorsAndrews NL, Lidke KA, Pfeiffer JR, Burns AR, Wilson BS, Oliver JM, Lidke DS,
JournalNat Cell Biol
PubMed ID18641640
The actin cytoskeleton has been implicated in restricting diffusion of plasma membrane components. Here, simultaneous observations of quantum dot-labelled FcepsilonRI motion and GFP-tagged actin dynamics provide direct evidence that actin filament bundles define micron-sized domains that confine mobile receptors. Dynamic reorganization of actin structures occurs over seconds, making the location ... More
Comparison of detection and signal amplification methods for DNA microarrays.
AuthorsVora GJ, Meador CE, Anderson GP, Taitt CR,
JournalMol Cell Probes
PubMed ID18675897
One of the factors limiting the use of DNA microarray technology for the detection of pathogenic organisms from clinical and environmental matrices has been inadequate assay sensitivity. To assess the effectiveness of post-hybridization secondary detection steps to enhance the sensitivity of DNA microarray-based pathogen detection, we evaluated a panel of ... More
Quantum dots based probes conjugated to annexin V for photostable apoptosis detection and imaging.
AuthorsLe Gac S, Vermes I, van den Berg A
JournalNano Lett
PubMed ID16967992
Quantum dots (Qdots) are nanoparticles exhibiting fluorescent properties that can be used for cell staining. We present here the development of quantum dots conjugated to Annexin V for specific targeting of apoptotic cells, for both apoptosis detection and staining of apoptotic "living" cells. For that purpose, Qdots Streptavidin Conjugates are ... More
Spectroscopic characterization of streptavidin functionalized quantum dots.
AuthorsWu Y, Lopez GP, Sklar LA, Buranda T
JournalAnal Biochem
PubMed ID17368555
The spectroscopic properties of quantum dots can be strongly influenced by the conditions of their synthesis. In this work, we have characterized several spectroscopic properties of commercial, streptavidin functionalized quantum dots (QD525, lot 1005-0045, and QD585, lot 0905-0031, from Invitrogen). This is the first step in the development of calibration ... More
Quantitative 3D fluorescence technique for the analysis of en face preparations of arterial walls using quantum dot nanocrystals and two-photon excitation laser scanning microscopy.
AuthorsFerrara DE, Weiss D, Carnell PH, Vito RP, Vega D, Gao X, Nie S, Taylor WR
JournalAm J Physiol Regul Integr Comp Physiol
PubMed ID16223849
Traditional imaging with one-photon confocal microscopy and organic fluorophores poses several challenges for the visualization of vascular tissue, including autofluorescence, fluorophore crosstalk, and photobleaching. We studied human coronary arteries (HCAs) and mouse aortas with a modified immunohistochemical (IHC) "en face" method using quantum dot (Qdot) bioconjugates and two-photon excitation laser ... More
Multispectral imaging of clinically relevant cellular targets in tonsil and lymphoid tissue using semiconductor quantum dots.
AuthorsFountaine TJ, Wincovitch SM, Geho DH, Garfield SH, Pittaluga S
JournalMod Pathol
PubMed ID16778828
Determination of the expression and spatial distribution of molecular epitopes, or antigens, in patient tissue specimens has substantially improved the pathologist's ability to classify disease processes. Certain disease pathophysiologies are marked by characteristic increased or decreased expression of developmentally controlled antigens, defined as Cluster of Differentiation markers, that currently form ... More