Detection of cell and tissue surface antigens using up-converting phosphors: a new reporter technology.
Identifieur interne : 000C74 ( Ncbi/Curation ); précédent : 000C73; suivant : 000C75Detection of cell and tissue surface antigens using up-converting phosphors: a new reporter technology.
Auteurs : H J Zijlmans [Pays-Bas] ; J. Bonnet ; J. Burton ; K. Kardos ; T. Vail ; R S Niedbala ; H J TankeSource :
- Analytical biochemistry [ 0003-2697 ] ; 1999.
Descripteurs français
- KwdFr :
- Animaux, Anticorps, Antigène spécifique de la prostate (analyse), Antigènes CD4 (analyse), Antigènes de surface (analyse), Avidine, Erbium, Humains, Immunohistochimie (), Lymphocytes (immunologie), Mesures de luminescence, Microscopie de fluorescence (), Microscopie de fluorescence (instrumentation), Mâle, Prostate (immunologie), Rayons infrarouges, Thulium, Yttrium.
- MESH :
- analyse : Antigène spécifique de la prostate, Antigènes CD4, Antigènes de surface.
- immunologie : Lymphocytes, Prostate.
- Animaux, Anticorps, Avidine, Erbium, Humains, Immunohistochimie, Mesures de luminescence, Microscopie de fluorescence, Mâle, Rayons infrarouges, Thulium, Yttrium.
English descriptors
- KwdEn :
- Animals, Antibodies, Antigens, CD4 (analysis), Antigens, Surface (analysis), Avidin, Erbium, Humans, Immunohistochemistry (methods), Infrared Rays, Luminescent Measurements, Lymphocytes (immunology), Male, Microscopy, Fluorescence (instrumentation), Microscopy, Fluorescence (methods), Prostate (immunology), Prostate-Specific Antigen (analysis), Thulium, Yttrium.
- MESH :
- chemical , analysis : Antigens, CD4, Antigens, Surface, Prostate-Specific Antigen.
- chemical : Antibodies, Avidin, Erbium, Thulium, Yttrium.
- immunology : Lymphocytes, Prostate.
- instrumentation : Microscopy, Fluorescence.
- methods : Immunohistochemistry, Microscopy, Fluorescence.
- Animals, Humans, Infrared Rays, Luminescent Measurements, Male.
Abstract
A novel luminescent reporter for the sensitive detection of antigens in tissue sections or on cell membranes is described. It consists of submicron-size phosphor crystals (0.2-0.4 microm), which are surface labeled with avidin or antibodies and capable of binding specifically to antigens on intact cells or in tissue sections. These phosphor reporters exhibit two-photon, anti-Stokes luminescence by up-converting infrared to visible light and are named Up-converting Phosphor Technology (UPT). They typically consist of yttriumoxysulfides doped with two different lanthanides exhibiting photostable, strong emission in the visible (blue, green, and red) upon excitation in the infrared. This report describes the conjugation of phosphor particles to NeutrAvidin with the subsequent use of this conjugate in a model system consisting of prostate-specific antigen in tissue sections and the CD4 membrane antigen on human lymphocytes. An epi-illumination fluorescence microscope was adapted to provide near-IR excitation using a xenon lamp for visualization of the visible emission. Advantages of UPT are (i) permanent, strong, anti-Stokes emission of discrete wavelengths; (ii) unmatched contrast in biological specimens due to the absence of autofluorescence upon excitation with IR light; (iii) simultaneous detection of multiple target analytes; and (iv) low-cost microscope modifications. The new methodology has not only high potential value in diagnostic pathology as described here, but may offer advantages for the detection of proteins or nucleic acids when applied to molecular biology, genomic research, virology, and microbiology.
DOI: 10.1006/abio.1998.2965
PubMed: 9918652
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pubmed:9918652Le document en format XML
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<profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals</term>
<term>Antibodies</term>
<term>Antigens, CD4 (analysis)</term>
<term>Antigens, Surface (analysis)</term>
<term>Avidin</term>
<term>Erbium</term>
<term>Humans</term>
<term>Immunohistochemistry (methods)</term>
<term>Infrared Rays</term>
<term>Luminescent Measurements</term>
<term>Lymphocytes (immunology)</term>
<term>Male</term>
<term>Microscopy, Fluorescence (instrumentation)</term>
<term>Microscopy, Fluorescence (methods)</term>
<term>Prostate (immunology)</term>
<term>Prostate-Specific Antigen (analysis)</term>
<term>Thulium</term>
<term>Yttrium</term>
</keywords>
<keywords scheme="KwdFr" xml:lang="fr"><term>Animaux</term>
<term>Anticorps</term>
<term>Antigène spécifique de la prostate (analyse)</term>
<term>Antigènes CD4 (analyse)</term>
<term>Antigènes de surface (analyse)</term>
<term>Avidine</term>
<term>Erbium</term>
<term>Humains</term>
<term>Immunohistochimie ()</term>
<term>Lymphocytes (immunologie)</term>
<term>Mesures de luminescence</term>
<term>Microscopie de fluorescence ()</term>
<term>Microscopie de fluorescence (instrumentation)</term>
<term>Mâle</term>
<term>Prostate (immunologie)</term>
<term>Rayons infrarouges</term>
<term>Thulium</term>
<term>Yttrium</term>
</keywords>
<keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Antigens, CD4</term>
<term>Antigens, Surface</term>
<term>Prostate-Specific Antigen</term>
</keywords>
<keywords scheme="MESH" type="chemical" xml:lang="en"><term>Antibodies</term>
<term>Avidin</term>
<term>Erbium</term>
<term>Thulium</term>
<term>Yttrium</term>
</keywords>
<keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Antigène spécifique de la prostate</term>
<term>Antigènes CD4</term>
<term>Antigènes de surface</term>
</keywords>
<keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Lymphocytes</term>
<term>Prostate</term>
</keywords>
<keywords scheme="MESH" qualifier="immunology" xml:lang="en"><term>Lymphocytes</term>
<term>Prostate</term>
</keywords>
<keywords scheme="MESH" qualifier="instrumentation" xml:lang="en"><term>Microscopy, Fluorescence</term>
</keywords>
<keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Immunohistochemistry</term>
<term>Microscopy, Fluorescence</term>
</keywords>
<keywords scheme="MESH" xml:lang="en"><term>Animals</term>
<term>Humans</term>
<term>Infrared Rays</term>
<term>Luminescent Measurements</term>
<term>Male</term>
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<keywords scheme="MESH" xml:lang="fr"><term>Animaux</term>
<term>Anticorps</term>
<term>Avidine</term>
<term>Erbium</term>
<term>Humains</term>
<term>Immunohistochimie</term>
<term>Mesures de luminescence</term>
<term>Microscopie de fluorescence</term>
<term>Mâle</term>
<term>Rayons infrarouges</term>
<term>Thulium</term>
<term>Yttrium</term>
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<front><div type="abstract" xml:lang="en">A novel luminescent reporter for the sensitive detection of antigens in tissue sections or on cell membranes is described. It consists of submicron-size phosphor crystals (0.2-0.4 microm), which are surface labeled with avidin or antibodies and capable of binding specifically to antigens on intact cells or in tissue sections. These phosphor reporters exhibit two-photon, anti-Stokes luminescence by up-converting infrared to visible light and are named Up-converting Phosphor Technology (UPT). They typically consist of yttriumoxysulfides doped with two different lanthanides exhibiting photostable, strong emission in the visible (blue, green, and red) upon excitation in the infrared. This report describes the conjugation of phosphor particles to NeutrAvidin with the subsequent use of this conjugate in a model system consisting of prostate-specific antigen in tissue sections and the CD4 membrane antigen on human lymphocytes. An epi-illumination fluorescence microscope was adapted to provide near-IR excitation using a xenon lamp for visualization of the visible emission. Advantages of UPT are (i) permanent, strong, anti-Stokes emission of discrete wavelengths; (ii) unmatched contrast in biological specimens due to the absence of autofluorescence upon excitation with IR light; (iii) simultaneous detection of multiple target analytes; and (iv) low-cost microscope modifications. The new methodology has not only high potential value in diagnostic pathology as described here, but may offer advantages for the detection of proteins or nucleic acids when applied to molecular biology, genomic research, virology, and microbiology.</div>
</front>
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