CAJ | 학술논문

报道了一种能用于活细胞双光子荧光( TPF)成像的水溶性聚合物纳米粒。首先以含双羧基的萘酰亚胺作为交联剂和荧光标记试剂，通过对聚乙烯亚胺发生交联反应制备纳米粒子，然后对其结构形态、单双光子荧光性能及细胞毒性进行测试。结果表明，获得的纳米粒为球形，粒径为5~10 nm；以443或800 nm为激发波长，荧光发射波长均为536 nm；在pH 4.0~9.0范围内，其荧光无明显变化；在pH 7.4的溶液中和激发光为443 nm 的条件下，对其连续测定1.2万次后荧光强度变化不超过1%，说明其酸碱稳定性和光稳定性较好；浓度在15 mg/L以下及与细胞作用时间在24 h以内细胞毒性较低。最后，用双光子共聚焦荧光显微镜观察了其在 Hela 细胞中的 TPF 成像性能。将Hela细胞与纳米粒共同孵育2 h后，在800 nm激光激发下，在细胞中可观察到其绿色荧光。此纳米粒可望用于靶向性双光子荧光成像探针的开发。
보도료일충능용우활세포쌍광자형광( TPF)성상적수용성취합물납미립。수선이함쌍최기적내선아알작위교련제화형광표기시제，통과대취을희아알발생교련반응제비납미입자，연후대기결구형태、단쌍광자형광성능급세포독성진행측시。결과표명，획득적납미립위구형，립경위5~10 nm；이443혹800 nm위격발파장，형광발사파장균위536 nm；재pH 4.0~9.0범위내，기형광무명현변화；재pH 7.4적용액중화격발광위443 nm 적조건하，대기련속측정1.2만차후형광강도변화불초과1%，설명기산감은정성화광은정성교호；농도재15 mg/L이하급여세포작용시간재24 h이내세포독성교저。최후，용쌍광자공취초형광현미경관찰료기재 Hela 세포중적 TPF 성상성능。장Hela세포여납미립공동부육2 h후，재800 nm격광격발하，재세포중가관찰도기록색형광。차납미립가망용우파향성쌍광자형광성상탐침적개발。
A water-soluble polymeric fluorescent nanoparticle ( PTCN ) has been proposed for two-photon fluorescence ( TPF ) imaging of living cells. PTCN was firstly prepared by chemical crosslinking of polyethyleneimine using a dual carboxyl 1, 8-naphthalimide derivative as the crosslinker. Subsequently, the investigations on its structure, morphology, one-and two-photon fluorescence properties and cytotoxicity were carried out. It was found that PTCN was spherical with the diameters of 5-10 nm. It could emit fluorescence at 536 nm with the excitation wavelengths of 443 nm and 800 nm, respectively. PTCN possessed good pH stability and photostability. Its fluorescence did not change in the pH range of 4. 0-9. 0. In pH 7. 4 buffer solution, when it was detected at λex/λem=443 nm/536 nm for 12000 times, the decrease in fluorescence intensity was less than 1%. Cytotoxic assays indicated it was low toxic upon interactions with Hela cells for 24 h at a concentration of 15. 0 mg/L. Finally, the application of PTCN for imaging live cells was assessed by TPF microscopy technique. After Hela cells were incubated with PTCN for 2 h, the green fluorescence was observed in the cellsap at the excitation wavelength of 800 nm. The proposed nanoparticle is potential in the development of targeting TPF probes.