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

1.瑞利光散射成像系统的建立及其在单个生物细胞中的应用用新研制的暗场显微镜系统对含金纳米颗粒的PBS缓冲液孵育前后的小鼠NIH3T3细胞进行了测量。通过扫描聚焦的白光获得散射图像，并获取其在各个点的光谱。纳米颗粒有时相互聚集，不均匀地分布在活细胞上。对纳米粒子的光谱也进行了详细的研究。用飞秒激光将纳米粒子注入单个生物细胞，并将其注入的DNAHE激光脉冲的表达聚焦到小鼠NIH3T3单细胞外，并将细胞上的聚合物纳米粒子引入细胞内。纳米粒子在细胞内的分布是局部的，在空间上是可控的。将增强型绿色荧光蛋白(EGFP)及其模型化合物(FITC Dextram)DNA质粒通过细胞膜和核膜直接照射注射。成功地表达了DNA，并对其条件进行了检测。飞秒激光照射后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照射与绿色荧光蛋白结合的小鼠NIH3T3细胞。荧光成像观察肌动蛋白应激纤维、细胞膜、丝状伪足和片状脂肪的损伤和修复过程。

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.