| 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.
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