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Titolo:
Combined laser tweezers and dielectric field cage for the analysis of receptor-ligand interactions on single cells
Autore:
Reichle, C; Sparbier, K; Muller, T; Schnelle, T; Walden, P; Fuhr, G;
Indirizzi:
Humboldt Univ, Lehrstuhl Membranphysiol, Inst Biol, Dept Biol Biophys, D-10115 Berlin, Germany Humboldt Univ Berlin Germany D-10115 ol Biophys, D-10115 Berlin, Germany Humboldt Univ, Charite, D-10115 Berlin, Germany Humboldt Univ Berlin Germany D-10115 v, Charite, D-10115 Berlin, Germany Humboldt Univ, Evotec Biosyst AG, D-10115 Berlin, Germany Humboldt Univ Berlin Germany D-10115 Biosyst AG, D-10115 Berlin, Germany
Titolo Testata:
ELECTROPHORESIS
fascicolo: 2, volume: 22, anno: 2001,
pagine: 272 - 282
SICI:
0173-0835(200101)22:2<272:CLTADF>2.0.ZU;2-B
Fonte:
ISI
Lingua:
ENG
Soggetto:
MICROELECTRODE SYSTEM; FORCE MEASUREMENTS; KINESIN MOLECULES; OPTICAL TWEEZERS; ADHESION; MANIPULATION; PARTICLES; BINDING; MECHANICS; SURFACES;
Keywords:
dielectric field cage; laser tweezers; adhesion; biotin-streptavidin; microsystem; dynamic force spectroscopy;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
38
Recensione:
Indirizzi per estratti:
Indirizzo: Fuhr, G Humboldt Univ, Lehrstuhl Membranphysiol, Inst Biol, Dept Biol Biophys, Invalidenstr 42, D-10115 Berlin, Germany Humboldt Univ Invalidenstr 42Berlin Germany D-10115 lin, Germany
Citazione:
C. Reichle et al., "Combined laser tweezers and dielectric field cage for the analysis of receptor-ligand interactions on single cells", ELECTROPHOR, 22(2), 2001, pp. 272-282

Abstract

A new technique based on the combination of optical and chip-based dielectrophoretical trapping was developed and employed to manipulate cells and beads with micrometer precision. The beads were trapped with optical tweezers(OT) and brought into contact for defined times with cells held in the dielectrophoretic field cage (DFC). The well-defined ligand-receptor system biotin-streptavidin was used to study the multiple interaction between biotinylated live cells and streptavidin-coated beads. The biotin density on the cell surface was varied down to a few single bonds (3 +/- 2 bonds/mum(2)) to control the valency of the binding. The quantitative relationship betweenthe contact area, ligand density and its diffusion rate in the outer membrane of the cell could be demonstrated. The increase of the strength of the cell-bead adhesion was strictly dependent on the increase of individual bond numbers in the contact area. This is in part due to accumulation of ligands (D similar to (0.5 +/- 0.1) 10(-8) cm(2)/s) in the contact area as seen by confocal laser scanning microscopy. Individual receptor-ligand rupture forces were evaluated and are compatible with values obtained by biomembraneforce probe techniques, To summarize, the combination leads to a new powerful microsystem for cell handling and pN-force measurements on the single-cell level.

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Documento generato il 04/07/20 alle ore 16:54:17