【作者】 李涛；

【导师】 唐宏武；

【作者基本信息】 武汉大学 ， 分析化学， 2004， 硕士

【摘要】 单细胞成像分析是细胞生物学和分析科学等领域的研究热点之一，其研究重点逐渐从细胞表面形态观测过渡到细胞内部亚细胞物质(细胞质、细胞膜及线粒体等)、单分子及单原子成分的结构特征、组分含量和生理功能等分析研究，因此发展高分辨和高灵敏度的单细胞成像技术已成为一项十分诱人的研究课题。本研究在前期工作基础上，发展了一种新型高分辨、高信噪比的成像分析方法—阿达玛变换(HT)显微成像术，通过采用归一化的HT图像，并将该系统成功应用于单细胞DNA倍性分析，并籍此对人乳腺肿瘤的良恶性进行判断；该系统还被用来和微流控单细胞进样技术联用，以实现动态单细胞定量分析。 本文工作主要集中在以下三个方面： 1．在前期工作基础上，对原HT显微镜装置的光路进行优化，使系统的分辨能力进一步提高；添加软件功能模块以获取归一化HT图像，并建立了一种在给定的标准下恢复图像的单细胞定量分析方法。 2．将HT显微镜成功应用于对单个癌细胞DNA倍性的测定和对肿瘤良恶性的判断，对26例人乳腺肿瘤的分析结果与病理学诊断很高的一致性。 3．将HT显微镜与微流控分析技术联用，发展了一种单细胞进样与在线成像分析新技术，并将其应用于花粉单细胞成像分析方面。结果表明，该方法结合了HT显微镜高分辨成像的性能和芯片高效进样的优点，因此能应用于单细胞分析领域，并能获得令人满意的结果。更多还原

【Abstract】 Single-cell imaging analysis is an advanced field in cell biology and analytical science. Its focus transfers gradually from observing cellular surface to studying the structure characteristic, components’ content and physiological functions of objects in cells such as sub-cell substance (cytoplasm, cell membrane and mitochondria), single molecule and single atom. Accordingly, the development of single-cell imaging techniques with high-resolution and high sensitivity is a fascinating research project. Based on our previous studies, this thesis presents a novel Hadamard transform(HT) microscopy with high resolution and high signal-to-noise, and by using normalized HT images the technique is applied to evaluate tumor malignancy based on the measurements of cellular DNA ploidy, and for a further study the technique is connected with the microfluidic single-cell sampling for dynamic cellular quantitative analysis.The contents of this thesis are three aspects as following:1. Based on the previous researches, the optical path of the prototype device of HT microscope is optimized to improve the resolving ability of the system. Some useful software is added to obtain normalized HT images, and a method for cellular quantitative analysis based on a given criterion for image recovery is developed.2. The HT microscopy is successfully used to measure DNA ploidy in single cancer cells for malignancy evaluation of tumors, and the analytical results of 26 cases of human breast tumors are highly in accordant with pathological diagnosis.3. By combining the HT microscopy with the microfluidic analysis technique, a novel method for single-cell sampling and on-line imaging is developed. This method is applied to single-cell imaging analysis of pollen cells. The result shows that it integrates the capability of high-resolution imaging of the HT microscopy and the advantage of high-efficiency sampling of the microchip, and thus can be applied in the field of single-cell analysis with satisfactory results.更多还原