小型化流式细胞仪的研制

发布时间：2018-01-06 06:27

本文关键词：小型化流式细胞仪的研制　出处：《浙江大学》2017年硕士论文　论文类型：学位论文

【摘要】：流式细胞仪是一种用于单个细胞多参数快速定量分析和分选的先进仪器,在基础科学研究与临床诊断中发挥着重要作用。然而,传统流式细胞仪体积庞大,结构精密,操作复杂,主要面向专业的科研型分析需求,并不适合场所流动性强的应用型分析需求。因此,基于微流控技术的流式细胞仪的小型化、集成化、便携化研究正逐渐成为该领域发展的一大主流方向。本论文主要包括以下三个章节的内容：第一章为文献综述,对流式细胞仪的原理概念、历史发展、结构组成等方面进行了全面的概括。进而,针对流式细胞仪中流体操控的核心——流体聚焦技术进行了具体讨论。最后,分类介绍了当前流式细胞仪在各领域中的广泛应用。第二章介绍了一台小型化单荧光流式细胞仪的研制工作。该仪器基于共焦型激光诱导荧光检测结构,以450 nm激光二极管为光源,实现前向散射光,侧向散射光以及绿色荧光的同步检测。利用3D打印技术设计制作了仪器的支撑结构,并以模块化的方式将各部分组合在一起,整体尺寸仅为26×6×12 cm3。仪器中使用了多个微型平移台结构,使部分模块可以实现小范围的微调,以获得最佳检测效果。液流部分采用十字套管结构,通过传统的流体力学聚焦模式实现对样品流的3D聚焦。在本章中,我们利用标准尺寸的绿色荧光微球对仪器检测性能进行了表征,并将其用于抗癌药物诱导的癌细胞凋亡检测,获得较理想的结果。第三章在第二章工作的基础上,初步进行了可实现多荧光检测的小型化流式细胞仪的研制工作。仪器采用共焦光路作为荧光检测结构,目标实现最多4色荧光,6参数的同步检测功能。目前,我们完成了液流部分结构的优化以及对侧向散射光和荧光检测光路中透镜位置的协调和优化。通过对套管结构中各管路位置进行调整,获得了更加稳定的且位置固定的3D鞘流。
[Abstract]:Flow cytometry is a kind of advanced instrument for rapid quantitative analysis of multi parameter and single cell sorting, plays an important role in basic scientific research and clinical diagnosis. However, the traditional flow cytometry in large volume, precise structure, complex operation, mainly for professional research demand analysis, is not suitable for application places the analysis of liquidity demand. Therefore, miniaturization, flow cytometry microfluidic technology based on integrated research, portable and has gradually become a mainstream direction of the development of this field. This paper mainly includes the following three chapters: the first chapter is the literature review, the concept of flow cytometry principle. The history of the development, structure composition were summarized. Then, aiming at the core flow cytometry in the control of fluid focusing technique were discussed in detail. Finally, the classification is introduced when Before the application of flow cytometry in various fields. The second chapter introduces the development of a miniature single fluorescence flow cytometry. The instrument structure of confocal laser induced fluorescence detection based on 450 nm laser diode as the light source, the light scattering, scattering light green fluorescence and synchronous detection the supporting structure of the instrument. Making use of 3D printing technology design, and in a modular way put all the pieces together, the overall size is only a plurality of micro displacement platform structure using 26 x 6 x 12 cm3. instruments, the module can achieve a small range of fine-tuning, to get the best result. The liquid flow part of the cross structure of casing, sample flow 3D focus by hydrodynamic focusing the traditional model. In this chapter, we use the green fluorescent microspheres standard size on detection performance were investigated, and For the detection of cancer cell apoptosis induced by anticancer drugs, obtained the ideal result. The third chapter in the second chapter, the preliminary research work can realize the miniaturization of multi fluorescence detection by flow cytometry. The instrument adopts the confocal optical path as fluorescence detection structure, the realization of the goal of a maximum of 4 color fluorescence 6, synchronous detection function parameters. At present, we have completed the flow optimization part of structure and the lateral scattering light and fluorescence detection of the position of the lens in the path of coordination and optimization. By adjusting the position of each pipeline casing structure, the more stable and fixed position of 3D sheath flow.

【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：Q-337

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