Multiplexed Elispot assay.
J Immunol Methods
; 341(1-2): 127-34, 2009 Feb 28.
Article
en En
| MEDLINE
| ID: mdl-19084532
ABSTRACT
Micron scale latex beads are well established as highly biocompatible reagents. Imbibing two fluorescent dyes into the interior of the beads enables the creation of a family of combinatorially colored labels. Previous use of such beads, in flow cytometry for example, has focused on beads of approximately 5 microm diameter. We show here that 280 nm combinatorially labeled particles can be used to create ELISA-style assays in 200 microm scale virtual wells, using digital microscopy as the readout. The utility of this technique is illustrated by profiling the secreted cytokine footprints of peripheral blood mononuclear cells in a multiparametric version of the popular Elispot assay. Doing so reveals noncanonical classes of T lymphocytes. We further show that the secreting cell type can be concurrently identified by surface staining with a cell type specific antibody conjugated to the same multiplexed beads.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Inmunoensayo
/
Linfocitos T
/
Citometría de Flujo
/
Microesferas
/
Anticuerpos
Límite:
Humans
Idioma:
En
Revista:
J Immunol Methods
Año:
2009
Tipo del documento:
Article
País de afiliación:
Estados Unidos