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Titolo:
Analysis of single-cell cultures by immunoaffinity capillary electrophoresis with laser-induced fluorescence detection
Autore:
Phillips, TM;
Indirizzi:
George Washington Univ, Med Ctr, Immunochem Lab, Washington, DC 20037 USA George Washington Univ Washington DC USA 20037 , Washington, DC 20037 USA
Titolo Testata:
LUMINESCENCE
fascicolo: 2, volume: 16, anno: 2001,
pagine: 145 - 152
SICI:
1522-7235(200103/04)16:2<145:AOSCBI>2.0.ZU;2-I
Fonte:
ISI
Lingua:
ENG
Soggetto:
INFLAMMATORY-BOWEL-DISEASE; NEUROPEPTIDES SUBSTANCE-P; GENE-RELATED PEPTIDE; LYMPHOCYTE-PROLIFERATION; PRECOLUMN DERIVATIZATION; BODY-FLUIDS; MICRODIALYSIS; SECRETION; RECEPTOR; ONLINE;
Keywords:
microdialysis; immunoaffinity capillary electrophoresis; laser-induced fluorescence detection;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
48
Recensione:
Indirizzi per estratti:
Indirizzo: Phillips, TM NIH, UAIR, DBEPS, ORS,OD, Bldg 13,Room 3N17,9000 Rockville Pike, Bethesda,MD 20892 USA NIH Bldg 13,Room 3N17,9000 Rockville Pike Bethesda MD USA 20892
Citazione:
T.M. Phillips, "Analysis of single-cell cultures by immunoaffinity capillary electrophoresis with laser-induced fluorescence detection", LUMINESCENC, 16(2), 2001, pp. 145-152

Abstract

Neuropeptide regulation of immunological activity is becoming an importantissue in both basic and clinical sciences, necessitating the need for analysis to be performed at the single-cell level. A microsampling procedure has been developed for studying secretion of biologically important peptides from neuropeptide-stimulated lymphocytes, based on microdialysis sampling coupled to immunoaffinity capillary electrophoresis (ICE), with laser-induced fluorescence (LIF) detection using a fibre-optic spectrometer and diode laser excitation. The system demonstrated a limit of detection in the high attomole (10(-18) mol/L) range with pure standards and was capable of monitoring secretion from a single cell over time. Using this system it was possible to differentiate the effects of four neuropeptides on both T and B cellrelease of regulatory cytokines. CB4(+) lymphocytes demonstrated a 7.5-fold increase in cytokine secretion over baseline following stimulation with substance P (SP) and calcitonin gene-related peptide (CGRP). B cells responded to CGRP and vasoactive intestinal peptide (VIP) stimulation (5.5-fold increase), but not to SP. These changes took place 12-20 h post-stimulation and, once the peak secretion had been reached, remained at that level for the duration of the experiment, This system demonstrates the ability to perform high sensitivity measurements on microsamples of biological fluids. Copyright (C) 2001 John Wiley & Sons, Ltd.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 03/04/20 alle ore 06:03:09