A sensor for adenosine triphosphate fabricated by laser-induced forward transfer of luciferase onto a poly(dimethylsiloxane) microchip
A sensor for adenosine triphosphate fabricated by laser-induced forward transfer of luciferase onto a poly(dimethylsiloxane) microchip
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DOI：
10.1016/j.apsusc.2007.04.013
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发表时间：
2007-08
影响因子：
6.7
通讯作者：
Yasuyuki Tsuboi;Yosuke Furuhata;N. Kitamura
中科院分区：
文献类型：
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作者：
Yasuyuki Tsuboi;Yosuke Furuhata;N. Kitamura
研究了激光诱导荧光素酶的正向转移（LIFT）作为一种潜在的技术，可用于制造微芯片三磷酸腺苷（ATP）传感器。选择聚二甲基硅氧烷（PDMS）作为荧光素酶沉积的底物。与其他固体底物（如玻璃和聚苯乙烯）相比，PDMS的柔韧性使其成为固定化荧光素酶微点的优越底物。利用355nm激光将荧光素酶成功地转移到PDMS底物上，同时保持了酶的生物活性。由于荧光素和ATP之间的酶促反应，从PDMS芯片上的一个转移点观察到归因于荧光素酶的黄色发光。通过将一个小光电二极管连接到PDMS芯片上，还制作了一个微芯片ATP传感器。在制备的微芯片的基础上，证实了光斑发光强度与ATP浓度之间的Michaelis-Menten关系。利用LIFT技术和PDMS衬底的组合制造生物传感器的潜力被证明是非常好的。
Laser-induced forward transfer (LIFT) of the enzyme luciferase was explored as a potential technique to be used in the fabrication of a microchip adenosine triphosphate (ATP) sensor. Poly(dimethylsiloxane) (PDMS) was selected as the substrate for deposition of the luciferase. In comparison with other solid substrates, such as glass and polystyrene, it was found that the flexibility of PDMS made it a superior substrate for the immobilization of micro-spots of luciferase. LIFT of luciferase onto a PDMS substrate using a 355nm laser was successfully carried out, while the bioactivity of the enzyme was maintained. Yellow luminescence ascribed to luciferase was observed from a transferred spot on the PDMS chip from the enzymatic reaction between luciferin and ATP. A microchip ATP sensor was also fabricated by attaching a small photodiode to the PDMS chip. On the basis of the fabricated microchip, the Michaelis–Menten relation between the luminescence intensity from the spot, and the ATP concentration was confirmed. The potential for fabricating biosensors using a combination of the LIFT technique with a PDMS substrate was shown to be very good.