Integration of multianalyte sensing functions on a capillary-assembled microchip: simultaneous determination of ion concentrations and enzymatic activities by a "drop-and-sip" technique.
A general and simple implementation of simultaneous multiparametric sensing in a single microchip is presented by using a capillary-assembled microchip (CAs-CHIP) integrated with the plural different reagent-release capillaries (RRCs), acting as various biochemical sensors. A novel "drop-and-sip" technique of fluid handling is performed with a microliter droplet of a model sample solution containing proteases (trypsin, chymotrypsin, thrombin, elastase) and divalent cations (Ca2+, Zn2+, Mg2+) that passes through the microchannel with the aid of a micropipette as a vacuum pump, concurrently filling each RRC via capillary force. To avert the evaporation of the nanoliter sample volume in each capillary, PDMS oil is dropped on the outlet hole of the CAs-CHIP exploiting the capillary force that results in spontaneous sealing of all the RRCs. In addition, this high-speed sample introduction alleviates the possibility of protein adsorption and capillary cross-contamination, allowing a reliable and multianalyte determination of a sample containing many different proteases and divalent cations by using the fluorescence image analysis. Presented results suggested the possible application of this microchip in the field of drug discovery and systems biology.
Henares
Terence G
TG
Graduate School of Material Science, University of Hyogo, 3-2-1 Kouto, Kamigori-cho, Ako-gun, Hyogo 678-1297, Japan.
Takaishi
Masayuki
M
Yoshida
Naoya
N
Terabe
Shigeru
S
Mizutani
Fumio
F
Sekizawa
Ryuichi
R
Hisamoto
Hideaki
H
D015374:Biosensing Techniques / Q000379:methods; D002413:Cations, Divalent / Q000032:analysis*; D046209:Electrophoresis, Microchip / Q000295:instrumentation / Q000379:methods*; D004798:Enzymes / Q000378:metabolism; D004867:Equipment Design; D005453:Fluorescence; D007477:Ions / Q000032:analysis; D010447:Peptide Hydrolases / Q000378:metabolism*
D002413:Cations, Divalent; D004798:Enzymes; D007477:Ions; D010447:Peptide Hydrolases