Creation of Reaction Fields in Single Bio Cells and Measurement of Their Reactions

1.瑞利光散射光谱/成像系统的研制及其在单细胞研究中的应用利用一种新研制的带暗视野显微镜的瑞利光散射光谱/成像系统，对单个小鼠NIH 3 T3细胞在含金纳米颗粒的PBS缓冲液中孵育前后的光散射特性进行了测量。通过扫描聚焦的白色光获得散射图像，并获得其在每个点处的光谱。纳米颗粒有时彼此聚集并且不均匀地分布在活细胞上。纳米粒子的光谱也进行了详细的检查。飞秒激光将纳米颗粒注入单个生物细胞及其注射DNA的表达激光脉冲聚焦在小鼠NIH 3 T3单个细胞外，并将细胞上的聚合物纳米颗粒引入其中。细胞内的纳米颗粒分布是局部的和空间可控的。通过直接照射细胞和核膜的方法，注射增强型绿色荧光蛋白（EGFP）及其模型化合物（FITC dextram）的DNA质粒。DNA的表达成功，并对其条件进行了检测.飞秒激光辐照生物细胞的结构变化及其荧光光谱成像研究用飞秒激光脉冲对肌动蛋白与绿色荧光蛋白（EGFP）结合的小鼠NIH 3 T3细胞进行成像和辐照。利用荧光成像技术对肌动蛋白应力纤维、细胞膜、丝状伪足和板状伪足的损伤和恢复过程进行了动态观察。

1. Development of Rayleigh light scattering spectroscopy/imaging system and its application to single bio cellsSingle mouse NIH3T3 cells incubated before and after a PBS buffer containing gold nanoparticles were measured by a newly developed system with a dark field microscope. The scattering images were obtained by scanning a focused white light, and its spectrum at each point was acquired. The nanoparticles are sometimes aggregated with each other and distributed inhomogeneously and on the living cells. Nanoparticle spectra were also examined in detail.2. Femtosecond laser injection of nanoparticles into single bio cells and their expression of injected DNAThe laser pulse was focused outside of mouse NIH3T3 single cells and the polymer nanoparticles on the cell were introduced into them. The nanosparticles distribution inside the cell is local and spatially controllable. DNA plasmid of enhanced green fluorescent protein (EGFP) and its model compound (FITC dextram) was injected by direct irradiation of cell and nucleus membranes. The expression of the DNA was successful and their conditions were examined.3. Structural changes of bio cells induced by femtosecond laser irradiation and their fluorescence spectral imagingMouse NIH3T3 cells whose actins are bound with EGFP were imaged and irradiated by femtosecond laser pulse. The dynamics of the resultant damages and recovery process of actin stress fiber, cell membrane, filopodia and lamellipodia were followed by fluorescence imaging.