Correlating short-term Ca(2+) responses with long-term protein expression after activation of single T cells.
Lab Chip
; 9(24): 3517-25, 2009 Dec 21.
Article
en En
| MEDLINE
| ID: mdl-20024031
In order to elucidate the dynamics of cellular processes that are induced in context with intercellular communication, defined events along the signal transduction cascade and subsequent activation steps have to be analyzed on the level of individual cells and correlated with each other. Here we present an approach that allows the initiation of cell-cell or cell-particle interactions and the analysis of cellular reactions within various regimes while the identity of each individual cell is preserved. It utilizes dielectrophoresis (DEP) and microfluidics in a lab-on-chip system. With high spatial and temporal precision we contacted single T cells with functionalized microbeads and monitored their immediate cytosolic Ca(2+) response. After this, the cells were released from the chip system and cultivated further. Expression of the activation marker molecule CD69 was analyzed the next day and correlated with the previously recorded Ca(2+) signal for each individual cell. We found a significant difference in the patterns of Ca(2+) traces between activated and non-activated cells, which shows that Ca(2+) signals in T cells can provide early information about a later reaction of the cell. Although T cells are non-excitable cells, we also observed irregular Ca(2+) transients upon exposure to the DEP field only. These Ca(2+) signals depended on exposure time, electric field strength and field frequency. By minimizing their occurrence rate, we could identify experimental conditions that caused the least interference with the physiology of the cell.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Activación de Linfocitos
/
Linfocitos T
/
Antígenos de Diferenciación de Linfocitos T
/
Antígenos CD
/
Regulación de la Expresión Génica
/
Calcio
Tipo de estudio:
Prognostic_studies
Límite:
Humans
Idioma:
En
Revista:
Lab Chip
Asunto de la revista:
BIOTECNOLOGIA
/
QUIMICA
Año:
2009
Tipo del documento:
Article
País de afiliación:
Alemania
Pais de publicación:
Reino Unido