Real-time monitoring of antibody secretion from B-cells on a microchip stimulated with a minute amount of mitogen

Abstract

This paper describes the cultivation of primary cells in a microchamber and the real-time monitoring of small amounts of antibody secretion after the introduction of a minute amount of stimulus by a microinjector using time-resolved fluorescence anisotropy analysis.

Isopentenyl pyrophosphate secreted from Zoledronate-stimulated myeloma cells, activates the chemotaxis of γδT cells

“…To investigate whether the supernatant of RPMI8226 MM cells treated with ZOL induced chemotaxis of gdT cells using a mTASbased microchamber [14,15], RPMI8226 MM cells were cultured with ZOL (1 mM) overnight. The conditioned medium (CM) was then collected and passed through a 0.22 mm filter (Millipore, Tokyo, Japan).…”

“…The microvalve is a particularly useful tool for investigating the fast-response of cells [10]. We have developed a micro total analysis system (mTAS)-based microfluidic cellular analysis device, which possesses a minute-volume (240 nL) chamber integrated with a microsample injector controlled by a microvalve that permits the injection of a small amount (several nL) of a solute [14,15]. We have previously demonstrated real-time monitoring of antibody secretion from B cells using this microchip [14].…”

“…We have developed a micro total analysis system (mTAS)-based microfluidic cellular analysis device, which possesses a minute-volume (240 nL) chamber integrated with a microsample injector controlled by a microvalve that permits the injection of a small amount (several nL) of a solute [14,15]. We have previously demonstrated real-time monitoring of antibody secretion from B cells using this microchip [14]. Moreover, because of the minute size of this chamber, we can analyze the behavior of individual cells in response to stimulators, including drugs and chemokines.…”

Towards real time analysis of protein secretion from single cells

“…14,15 The analytical challenges on single cell level, however, are small numbers of molecules that are metabolized and/or secreted by the cell of interest. While in most studies, addressing single to few cells, the accumulation of metabolites over time is monitored in a microchamber, 16,17 we aimed at the analysis of individual secreted molecules in a micro flow. Thus, time resolved changes in cell physiology can be determined, as signal accumulation is prevented by micro flow.…”