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Micropatterned Porous Silicon Photonic Crystals towards Microarray-based Cell Sensing

机译:微图案化多孔硅光子晶体向基于微阵列的细胞传感

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摘要

Porous silicon (PSi) is an ideal platform for label-free biosensing, and the development of PSi patterning will open a pathway to the development of highly parallel PSi biosensors. The primary aim of this research is to develop a PSi photonic crystal based microarray to monitor molecule secreted from cells by the optical reflectivity properties. In order to achieve the research aim, a number of works must be achieved: 1) to provide a reference for monitoring the analyte of interest from cell microarrays, real-time monitoring of the analyte of interest in multiple spots must be achieved. 2) A suitable microfabrication technique must be developed, so that different chemical functionalities can be introduced to discrete PSi surface and can be used to mediate special control of the cell adhesion to form microarrays. 3) The fabricated PSi microarrays must be applied to monitor and read out the cellular response from different numbers of cells and even single-cell level. In relation to the research aims, Chapter 3 presents the work of using PSi chip modified with gelatin to detect protease from both free protease solution and live cells. The monitoring is carried out continuously and read out from multiple locations over the surface. Chapter 4 introduces a photolithographic strategy for chemically patterning the PSi with Ti hard mask, and characterization methods including IR microscopy and reflectivity scanning are explored to obtain the spatial information from the patterned PSi chip. Chapter 5 presents a work based a photolithographic strategy with photoresist mask instead of Ti mask. Fluorescence labelled BSA and RGD peptide are conjugated to PSi surface to demonstrate the ability of patterning biological species. Detection of protease from different array elements on the patterned PSi chip was demonstrated by monitoring blue-shifts in the reflectivity resulted from gelatin cleavage by protease. In Chapter 6, cell microarray is achieved on PEG hydrogel patterned PSi surface using direct photopatterning method. Optical monitoring of live cells from the cell microarray is carried out by measuring the optical shift during cell osmotic lysis. A preliminary result of monitoring stimulated cells using the cell microarray is also carried out. Chapter 7 concludes the work with a discussion of lessons learned and the perspectives for future progress with PSi as a microarray platform for further biomedical applications.
机译:多孔硅(PSi)是无标签生物传感的理想平台,而PSi图案的开发将为高度平行的PSi生物传感器的开发开辟道路。这项研究的主要目的是开发一种基于PSi光子晶体的微阵列,以通过光反射特性来监测细胞分泌的分子。为了达到研究目的,必须完成许多工作:1)为监测细胞微阵列中的目标分析物提供参考,必须对多个点中的目标分析物进行实时监测。 2)必须开发一种合适的微加工技术,以便可以将不同的化学功能引入离散的PSi表面,并可以用于介导细胞粘附的特殊控制以形成微阵列。 3)必须将制造的PSi微阵列应用于监测和读出来自不同细胞数量甚至单细胞水平的细胞反应。关于研究目标,第3章介绍了使用明胶修饰的PSi芯片从游离蛋白酶溶液和活细胞中检测蛋白酶的工作。监视是连续进行的,并且可以从表面上的多个位置读出。第4章介绍了用Ti硬掩模对PSi进行化学构图的光刻策略,并探索了包括IR显微镜和反射率扫描在内的表征方法,以从构图的PSi芯片中获得空间信息。第5章介绍了基于工作原理的光刻策略,该工艺采用光致抗蚀剂掩模代替Ti掩模。荧光标记的BSA和RGD肽与PSi表面偶联,以证明图案化生物物种的能力。通过监测明胶被蛋白酶裂解产生的反射率的蓝移,证明了从图案化的PSi芯片上不同阵列元素中检测到蛋白酶。在第6章中,使用直接光图案化方法在PEG水凝胶图案化的PSi表面上实现了细胞微阵列。通过测量细胞渗透裂解过程中的光学位移,可以对细胞微阵列中的活细胞进行光学监控。还进行了使用细胞微阵列监测刺激细胞的初步结果。第7章在总结工作时总结了经验教训,并展望了PSi作为进一步生物医学应用的微阵列平台的未来进展。

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