Microfluidic Chip toward Cellular ATP and ATP-Conjugated Metabolic Analysis with Bioluminescence Detection

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• 作者：Bi-Feng Liu ; Motoaki Ozaki ; Hideaki Hisamoto ; Qingming Luo ; Yuichi Utsumi ; Tadashi Hattori ; and Shigeru Terabe
• 刊名：Analytical Chemistry
• 出版年：2005
• 出版时间：January 15, 2005
• 年：2005
• 卷：77
• 期：2
• 页码：573 - 578
• 全文大小：158K
• 年卷期：v.77,no.2(January 15, 2005)
• ISSN：1520-6882

文摘

In this article, a microfluidic platform integrating capillaryelectrophoresis and bioluminescence (BL) detection thatwas fabricated in poly(dimethylsiloxane) (PDMS) with lab-on-a-chip technology was demonstrated for cellular metabolic analyses. A microchannels network, "cross combining with Y", was designed to perform on-chip samplepreparation, separation, and BL detection of ATP andATP-conjugated metabolites, using firefly luciferin-luciferase BL system. A dynamic modification of the channelwall of PDMS proved to be crucial to reverse the directionof electroosmotic flow (EOF), which was uniquely achievedby a prewash cycle with a cationic surfactant didodecyldimethylammonium bromide. The influences of surfactant on the EOF and BL reaction were also investigated.Quantitative analyses revealed a dynamic linear rangeover 2 orders of magnitude for ATP, with a detection limitdown to submicromolar (midattomole). The method wasvalidated by measuring cellular ATP of E. coli. with directon-chip cell lysis. Further work was emphasized on ATP-conjugated metabolite analysis, using galactose as anexample. Assays of galactose in human urine samplesconfirmed the reliability of the protocol, which revealedgood prospect of this platform for ATP-conjugated submetabolomic profiling.