Differential measurement with a microfluidic device for the highly selective continuous measurement of histamine released from rat basophilic leukemia cells (RBL-2H3)

We fabricated a micro-fluidic device for the highly selective detection of the histamine released from rat basophilic leukemia (RBL) 2H3 cells. The device has two thin layer flow channels, each with one working electrode. One electrode was modified with Os-polyvinylpyridine based mediator containing horseradish peroxidase (Os-gel-HRP) and histamine oxidase (HAOx), the other was modified with Os-gel-HRP without any HAOx. We employed the device for differential measurement by using the HAOx modified electrode for detection and the unmodified electrode as a reference. The detection limit was greatly improved from 190 to 25 nM since the baseline noise level was suppressed. We used differential measurement to observe the histamine released from RBL-2H3 cells when stimulated with dinitrophenylated bovine serum albumin (DNP-BSA) as an antigen. We injected 5 muM of histamine solution into our device and it remained stable for more than 8 h.