Development of a microchip-based bioassay system using cultured cells.
We developed a novel bioassay system using a glass microchip and cultured cells. A microchamber for cell culture and microchannels for reactions and detection were fabricated on a Pyrex glass substrate by photolithography and wet etching techniques. Cell culture, chemical and enzymatic reactions, and detection were integrated into the microchip. To keep different temperatures locally in three areas of the microchip, we designed and fabricated a temperature control device. Nitric oxide released from macrophage-like cells stimulated by lipopolysaccharide was successfully monitored with the microchip, the temperature control device, and a thermal lens microscope. The total assay time was reduced from 24 to 4 h, and detection limit of NO was improved from 1 x 10(-6) to 7 x 10(-8) M compared with conventional methods. Moreover, the system could monitor a time course of the release, which is difficult to measure by conventional batch methods. We conclude that this system is promising for a rapid bioassay system with very small consumption of cells.
Goto
Makiko
M
Department of Applied Chemistry, School of Engineering, University of Tokyo, Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.
Sato
Kiichi
K
Murakami
Atsushi
A
Tokeshi
Manabu
M
Kitamori
Takehiko
T
D000818:Animals; D001681:Biological Assay / Q000379:methods*; D002138:Calibration; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D004867:Equipment Design; D056656:Lab-On-A-Chip Devices; D008070:Lipopolysaccharides / Q000494:pharmacology; D017737:Macrophages, Peritoneal / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D046208:Microchip Analytical Procedures / Q000379:methods*; D009569:Nitric Oxide / Q000032:analysis*; D012680:Sensitivity and Specificity; D013696:Temperature
D008070:Lipopolysaccharides; D009569:Nitric Oxide