创新的光学技术与单细胞分析方法，能够通过监测单个细胞生理变化的细节获知生命活动途经并阐释其功能和分子机制。家蚕微孢子虫(Nosema bombycis，简称N.b)是家蚕微粒子病的病原真菌, 是养蚕业的严重威胁。本项目基于单细胞拉曼光谱、多细胞相差显微成像和荧光显微成像等光学新技术，实时监测和分析多个N.b单孢子发芽过程海藻糖的变化及胞内其它生物大分子的实时变化动态信息，获知：（1）不同微孢子虫胞内物质的异质性；（2）N.b孢子发芽过程孢内主要生物大分子的变化动态；（3）物理、化学因子对N.b孢子发芽的影响及其分子机制；（4）N.b孢子发芽的异质性以及造成这种异质性的机理。项目通过创新技术和创新方法了解N.b孢子的发芽规律和异质性，为认识微孢子虫的发芽机制与侵染机制，也为阻断或降低微孢子虫对寄主的侵染提供科学依据和理论参考。同时，项目的预期成果也为光学技术应用于生物医学单细胞研究提供借鉴。

Novel optical techniques and single-cell analysis could help reveal the path of life activity, illuminate its functions and molecular mechanisms via detailed change of individual cells. Pebrine disease caused by infections, parasitic of Nosema (Nosema bombycis, N.b) is a traditional and destructive disease in sericulture. In this project, multi-focus laser tweezers Raman spectroscopy, phase contrast microscopy and fluorescence microscopy are used to analyze the real-time dynamic germination of hundreds of individual N.b spores, by monitoring the kinetics of intracellular trehalose and biomolecules of individual spores. This cross-disciplinary research project attempts to reveal: (i) the intracellular and intercellular heterogeneity of N.b spores. (ii) the kinetics of intracellular biomolecules of N.b spore germination in-vitro; (iii) the effect of physical and chemical agents on the N.b spores and spore germination，and its molecular mechanisms; and (iv) the heterogeneity of N.b spore germination and its mechanisms. The knowledge derived from this work will allow us to better understand the process of spore germination of N.b and its heterogeneity, and provide scientific help to develop new treatments to block or reduce the microsporidian infection of host. The results will also provide useful reference to single-cell analysis in biomedical field.