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Integration of Microsphere Resonators with Bioassay Fluidics for Whispering Gallery Mode Imaging

机译:微球谐振器与生物测定流体学的集成用于耳语画廊模式成像

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摘要

Whispering gallery mode resonators are small, radially symmetric dielectrics that trap light through continuous total internal reflection. The resonant condition at which light is efficiently confined within the structure is linked with refractive index, which has led to the development of sensitive label-free sensing schemes based on whispering gallery mode resonators. One resonator design uses inexpensive high index glass microspheres that offer intrinsically superior optical characteristics, but have proven difficult to multiplex and integrate with the fluidics for sample delivery and fluid exchange necessary for assay development. Recently, we introduced a fluorescence imaging approach that enables large scale multiplexing with microsphere resonators, thus removing one obstacle for assay development. Here we report an approach for microsphere immobilization that overcomes limitations arising from their integration with fluidic delivery. The approach is an adaptation of a calcium-assisted glass bonding method originally developed for microfluidic glass chip fabrication. Microspheres bonded to glass using this technique are shown to be stable with respect to fluid flow and show no detectable loss in optical performance. Measured Q-factors, for example, remain unchanged following sphere bonding to the substrate. The stability of the immobilized resonators is further demonstrated by transferring lipid films onto the immobilized spheres using the Langmuir-Blodgett technique. Bilayers of DOPC doped with GM1 were transferred onto immobilized resonators to detect the binding of cholera toxin to GM1. Binding curves generated from shifts in the whispering gallery mode resonance result in a measured Kd of 1.5 × 10−11 with a limit of detection of 3.3 pM. These results are discussed in terms of future assay development using microsphere resonators.
机译:回音壁模式谐振器是小型的,径向对称的电介质,可通过连续的全内反射来捕获光。有效地将光限制在结构内的共振条件与折射率有关,这导致了基于耳语画廊模式共振器的灵敏的无标记传感方案的发展。一种谐振器设计使用廉价的高折射率玻璃微球,这些玻璃微球本质上具有出色的光学特性,但事实证明很难与流体学进行多路复用和集成,以进行分析开发所需的样品传输和流体交换。最近,我们推出了一种荧光成像方法,可以与微球共振器进行大规模多路复用,从而消除了分析开发的障碍。在这里,我们报告了一种微球固定化的方法,该方法克服了由于它们与流体输送相结合而引起的局限性。该方法是最初为微流体玻璃芯片制造开发的钙辅助玻璃粘合方法的改编。使用这种技术键合到玻璃上的微球显示出对流体流动稳定,光学性能没有可检测到的损失。例如,在球体粘结到基板后,测得的Q因子保持不变。通过使用Langmuir-Blodgett技术将脂质膜转移到固定的球体上,进一步证明了固定的谐振器的稳定性。将掺有GM1的DOPC双层转移到固定的共振器上,以检测霍乱毒素与GM1的结合。从耳语回廊模式共振的移位产生的结合曲线导致测得的Kd为1.5×10 -11 ,检出限为3.3 pM。这些结果将在使用微球共振器的未来分析开发中进行讨论。

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