利用N-芴甲氧羰基-L-赖氨酸分子接枝异硫氰基荧光素,合成赖氨酸-荧光素小分子,在反相微乳液中京尼平作为交联剂交联赖氨酸-荧光素分子制备超小纳米粒。所得纳米粒纯化后进行紫外-近红外可见光扫描分析,确定荧光素交联到纳米粒上及未反应小分子被完全洗脱。动态光散射考察所得纳米粒径为(198.33±0.03)nm、透射电子显微镜观察到纳米粒呈球形,平均粒径为39.78nm。荧光光谱表征所得纳米粒与荧光素荧光光谱完全吻合。利用胶质瘤U87细胞进行纳米粒荧光成像,细胞被染色。利用CHO细胞考察纳米粒的毒性,在一定浓度下(小于8mg/mL)纳米粒对细胞无明显毒副作用。
Preparation of a kind of ultrasmall fluorescent lysine nanoparticle for Glioblastoma fluorescent ima-ging was as follows: first, the lysine-fluorescein (Lys-FITC) moleculewas synthesizedthrough fluorescein isothiocyanateand reaction with amino group of Fmoc-Lys-OH ·HC1; second, the end-product Lys-FITC was cross-linked by genipin in reversed microemulsion. After purification, UV absorbance spectrum was investiga-ted to validate no free Fluorescein isothiocyanate remained. Fluorescence property was measured on PerkinElmer LS 55 which showed fluorescence spectrum not changed. The particle size and morphology were observed by dynamic light scattering (DLS) and transmission electron microscope (TEM), which showed (198.33±0.03) nm and 39.78 respectively. Finally, Lys-FITC nanoparticle was cultured with U87 cell for fluo-rescent imaging and imaging signal in U87 cell was clearly visible. CHO cell tested by CCK-8 cultured with different concentration of nano confirmed no cytotoxicity when bellow 8 mg/mL.