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

1. 瑞利光散射光谱/成像系统的建立及其在单个生物细胞中的应用采用新研制的系统，在暗场显微镜下对含金纳米粒子的PBS缓冲液前后孵育的小鼠NIH3T3细胞进行了测量。通过扫描聚焦白光获得散射图像，并获取其在各点的光谱。纳米颗粒有时相互聚集，不均匀地分布在活细胞上。纳米粒子的光谱也进行了详细的研究。飞秒激光将纳米粒子注入单个生物细胞及所注射dna的表达。将激光脉冲聚焦于小鼠NIH3T3单细胞外，将聚合物纳米粒子注入细胞内。纳米粒子在细胞内的分布具有局部性和空间可控性。采用直接照射细胞和核膜的方法注射增强型绿色荧光蛋白（EGFP）的DNA质粒及其模型化合物（FITC dextram）。DNA表达成功，并对其条件进行了检测。飞秒激光辐照诱导生物细胞的结构变化及其荧光光谱成像用飞秒激光脉冲对动蛋白与EGFP结合的小鼠NIH3T3细胞进行成像和照射。荧光成像观察了肌动蛋白应激纤维、细胞膜、丝状足和板足的损伤动态和恢复过程。

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.