Functionalized nanocrystal-tagged fluorescent polymer beads: synthesis, physicochemical characterization, and immunolabeling application.
A methodology for incorporating solubilized CdSe/ZnS core/shell nanocrystals (NCs) into functionalized carboxylated polystyrene latexes 0.3-1 microm in diameter via a swelling procedure was developed and used for the production of homogeneous, highly fluorescent polymeric beads (HFPBs), which were found to be comparable in brightness to standard polymeric microspheres doped with organic fluorophores and more photostable than the latter by more than 50 times (Fluoresbrite yellow-orange microspheres were used as an example). The three-dimensional (3D) confocal analysis of individual 1-microm HFPB demonstrated that the beads were doped with the NCs almost homogeneously. HFPBs 0.3 microm in diameter were conjugated with anti-mouse polyvalent immunoglobulins and used for immunofluorescent detection of p-glycoprotein, a mediator of the multidrug resistance phenotype, overexpressed in the membrane of MCF7r breast adenocarcinoma cells. The photostability of NCs-tagged HFPBs offers obvious advantages for the reconstruction of 3D confocal fluorescence images of antigen distribution, and their exceptionally high brightness combined with photostability permits the detection of a single antigen molecule using a standard epifluorescence microscope.
Stsiapura
Vitali
V
EA no 3798 Détection et Approches Thérapeutiques Nanotechnologiques dans les Mécanismes Biologiques de Défense, Institut Fédératif de Recherche No. 53 Biomolécules, Université de Reims Champagne-Ardenne, 51 rue Cognacq Jay, 51100 Reims, France.
Sukhanova
Alyona
A
Artemyev
Mikhail
M
Pluot
Michel
M
Cohen
Jacques H M
JH
Baranov
Alexandre V
AV
Oleinikov
Vladimir
V
Nabiev
Igor
I
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1 / Q000235:genetics / Q000378:metabolism; D000906:Antibodies / Q000276:immunology*; D045744:Cell Line, Tumor; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D003460:Crystallization; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D008863:Microspheres; D049329:Nanostructures / Q000737:chemistry*; D011108:Polymers / Q000138:chemical synthesis* / Q000737:chemistry*; D012995:Solubility; D013057:Spectrum Analysis
D020168:ATP Binding Cassette Transporter, Subfamily B, Member 1; D000906:Antibodies; D005456:Fluorescent Dyes; D011108:Polymers