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
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
AuthorsSweeney E, Ward TH, Gray N, Womack C, Jayson G, Hughes A, Dive C, Byers R,
JournalBiochem Biophys Res Commun
PubMed ID18621021
'Quantum dots are photostable fluorescent semiconductor nanocrystals possessing wide excitation and bright narrow, symmetrical, emission spectra. These characteristics have engendered considerable interest in their application in multiplex immunohistochemistry for biomarker quantification and co-localisation in clinical samples. Robust quantitation allows biomarker validation, and there is growing need for multiplex staining due ... 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
Chronic spinal cord injury impairs primary antibody responses but spares existing humoral immunity in mice.
AuthorsOropallo MA, Held KS, Goenka R, Ahmad SA, O'Neill PJ, Steward O, Lane TE, Cancro MP,
JournalJ Immunol
PubMed ID22523388
Spinal cord injury (SCI) results in immune depression. To better understand how injury inhibits humoral immunity, the effects of chronic thoracic SCI on B cell development and immune responses to thymus-independent type 2 and thymus-dependent Ags were determined. Mice received complete crush injury or control laminectomy at either thoracic level ... More
Quantum dot semiconductor nanocrystals for immunophenotyping by polychromatic flow cytometry.
AuthorsChattopadhyay PK, Price DA, Harper TF, Betts MR, Yu J, Gostick E, Perfetto SP, Goepfert P, Koup RA, De Rosa SC, Bruchez MP, Roederer M
JournalNat Med
PubMed ID16862156
Immune responses arise from a wide variety of cells expressing unique combinations of multiple cell-surface proteins. Detailed characterization is hampered, however, by limitations in available probes and instrumentation. Here, we use the unique spectral properties of semiconductor nanocrystals (quantum dots) to extend the capabilities of polychromatic flow cytometry to resolve ... 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