口腔医学研究

口腔医学研究 ›› 2017, Vol. 33 ›› Issue (11): 1213-1217.DOI: 10.13701/j.cnki.kqyxyj.2017.11.020

• 其他研究 • 上一篇    下一篇

色氨酸碳点的合成及在细胞生物学成像中的应用

唐秋玲1, 赵晓欢2, 杨明锡2, 潘佳慧1, 李格格1, 孟阳1, 王柳然1, 于维先3*   

  1. 1. 吉林大学口腔医学院牙周病科 吉林 长春 130021;
    2. 吉林大学超分子结构与材料国家重点实验室 吉林 长春130012;
    3. 吉林省牙发育及颌骨重塑与再生重点实验室 吉林 长春 130021
  • 收稿日期:2017-05-10 出版日期:2017-11-20 发布日期:2017-11-29
  • 通讯作者: 于维先,电话:0431-88796000
  • 作者简介:唐秋玲(1990～ ),女,湖北人,医师,硕士,主要从事口腔牙周病学临床和基础研究工作。
  • 基金资助:
    国家自然科学基金面上项目(编号:81570983)
    吉林省卫生技术创新项目(编号:2016J073)
    吉林省科技厅自然科学基金项目(编号:20150101076JC)

Preparation and Cell Biology Imaging with Tryptophan Carbon Dots

Tang Qiu-Ling1, ZHAO Xiao-huan2, YANG Ming-xi2, PAN Jia-hui1, LI Ge-ge1, MENG Yang1, WANG Liu-ran1, YU Wei-xian3*   

  1. 1. Department of Periodontology, School of Stomatology, Jinin University. Changchun 130021, China;
    2. State Key Lab of Supramolecular Structure and Material, Jinin University. Changchun 130012, China;
    3. Key Lab for Tooth Development and Jaw Regeneration of Jinin Province. Changchun 130021, China
  • Received:2017-05-10 Online:2017-11-20 Published:2017-11-29

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343

被引次数

1

摘要： 目的:利用碳点标记不同细胞,通过共聚焦显微镜下碳点的多色荧光特性,达到观察不同细胞的成像特点。方法:采用水热法合成色氨酸碳点,用TEM、FT-IR、荧光光谱仪对碳点进行表征。采用MTT法检测碳点对小鼠单核巨噬细胞系RAW264.7、小鼠前成骨细胞系MC3T3-E1和小鼠成纤维细胞系L929成像效果,以确定碳点最佳成像浓度。选择最佳浓度的碳点分别与上述3种细胞共培养24 h后,在共聚焦显微镜下观察细胞的成像特点。结果:TEM下观察制备的碳点为球形颗粒,粒径约为3.35 nm;FT-IR显示碳点的分子结构仅由碳、氧、氢和氮元素组成。荧光光谱显示碳点在不同激发波长下可以呈现不同的颜色,具有激发依赖性。MTT结果显示当碳点浓度在400 mg/L时,RAW264.7细胞活性达67%,MC3T3-E1细胞活性达79%,L929细胞活性达89%,表明碳点进入细胞内部并不会明显影响细胞活性。成像结果显示,在488 nm和543 nm激发波长下,3种细胞均可以呈现绿色和红色的荧光图像,RAW264.7和L929荧光区主要集中在细胞膜和细胞质,而MC3T3-E1整个细胞都具有荧光,表明碳点可能部分进入细胞核中。结论:制备的碳点对细胞成像效果好,且能够使细胞具有多色荧光,对直接观察和分析细胞的形态和生理活动具有重要的意义。

关键词: 荧光碳点, 细胞, 生物学成像

Abstract: Objective: To investigate the imaging characteristics of different cells that labeled by carbon dots (CDs) with multicolor fluorescence properties. Methods: Tryptophan CDs were synthesized by hydrothermal method and characterized by TEM, FT-IR, and fluorescence spectrometer. MTT method was used to detect CDs at different concentrations co-cultured with RAW264.7, MC3T3-E1, and L929 for 24h, which were observed under confocal microscope. The optimum imaging concentration of CDs was determined. Results: TEM showed that CDs were spherical particles with excellent dispersion and particle size of about 3.72nm. FTIR showed that molecular structure of CDs was consisted of carbon, oxygen, hydrogen, and nitrogen elements. Fluorescence spectra showed that at different excitation wavelengths, CDs could present different colors with excitation dependencies. MTT results showed that the cell activity of RAW264.7, MC3T3-E1, and L929 were up to 67%, 79% and 89% when the concentration of CDs was 400mg/L. The results of cell imaging showed that green and red fluorescence images were observed at 488 nm and 543 nm excitation wavelengths. The fluorescence of CDs was mainly detected in cell membrane and cytoplasm of RAW264.7 and L929. The entire MC3T3-E1 cell showed fluorescence, which indicated that the CDs might partially enter into its nucleus. Conclusion: Tryptophan CDs can be applied to cell imaging to facilitate observation and analysis of cell morphology and physiological activities.

Key words: Carbon dots, Cell, Biological imaging

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