基于空芯光子晶体光纤载体的活细胞生物激光器

In June of 2011，《Nature Photonics》reported the first live biological laser built by two photomedical researach scientists in Harvard medical school.It immediately attracted huge attention from scientists and the media. It was considered a big breakthrough in the photonics and biological world. The team engineered human embryonic kidney cells to produce GFP, then placed a single cell between two mirrors to make a resonant cavity just 20 micron across. When they pumped the cell with pulses of blue laser, it emitted a directional green laser beam visible with the naked eye - and the cell was still alive. The birth of the first biological laser opened a new door for both biologists and photonics researchers. There is vast unknown information needs to be explored. It is in this context, we proposed our research project. In this project,we will engineer human breast cancer cells MCF7 to stably express green fluorescent protein (GFP), then utilize a hollow-core photonic crystal fiber as optical resonant cavity, fill the cavity with engineered MCF7 cells, and pump the cells with our home-made nanosecond pulsed blue laser. We expect that green laser would be emitted from the live MCF7 cells. Since we improved the resonant cavity design, more than one cell would be on the optical path. We anticipate that the laser output would be greater than the published single cell laser. In addition, our design would be easier to perform and more suitable for in vivo experiments. Our results together with published data will pave a new path for utilizing optical strategies to solve biomedical problems. It also holds great potential for future applications, such as light-based therapeutics, diagnosis for human diseases (such as cancer) and imaging, designing novel biological optical elements etc. Since the cells can regenerate, the biological laser or optical element would be self-healing and regenaratable. The Successful implementation of our project will display the first live cell biological laser in China, which will place China's biological laser research in the frontier of the world.

2011年6月，《Nature Photonics》报道了美国哈佛大学两位科学家研制成功世界首台活的单细胞生物激光器。他们将表达绿色荧光蛋白（GFP）的人类肾细胞置于由两片反射镜组成的高Q值谐振腔中，用脉冲蓝光泵浦，实现了活细胞绿色激光输出。这在激光发展史上是一极其重要的突破，为光子学在生命科学中的应用开辟了一个崭新的领域。然而，作为基础性前沿研究，其研究方法亟需进一步创新，科学问题亟待更加深入的探索。本申请拟将稳定表达GFP的人类乳腺癌细胞灌注于空芯光子晶体光纤中，利用自行研制的纳秒脉冲蓝光激光器对光纤中的细胞进行泵浦，以实现活细胞激光输出。原来的单细胞激光器将变为多细胞激光器，激光输出将会显著增强。此研究亦可使我们通过分析生物激光所携带的信息来研究细胞内部的生理过程。成功完成拟议的研究将首次展示以空芯光子晶体光纤为载体的多细胞生物激光器，使我国活细胞激光器的研究在世界上占有一席之地。

2011年6月，《Nature Photonics》报道了美国哈佛大学两位科学家研制成功世界首台活的单细胞生物激光器。他们将表达绿色荧光蛋白（GFP）的人类肾细胞置于由两片反射镜组成的高Q值谐振腔中，用脉冲蓝光泵浦，实现了活细胞绿色激光输出。这在激光发展史上是一极其重要的突破，为光子学在生命科学中的应用开辟了一个崭新的领域。然而，作为基础性前沿研究，其研究方法亟需进一步创新，科学问题亟待更加深入的探索。本项目综合利用激光光子学的研究手段和细胞生物学的研究方法，在研制小型化纳秒脉冲可见激光泵浦源的基础上，采用空芯光子晶体光纤为荧光蛋白溶液或活细胞载体，首先研制荧光蛋白溶液的激光器，在此基础上再研制具有荧光表达的活细胞生物激光器；对荧光表达的活细胞的荧光性能、激光性能、激光模式及抗激光损伤阈值等性能进行探索性研究，并观察分析活细胞在激光输出条件下的生物学特性，为活细胞生物激光器将来在光子学和生物活体中的应用打下坚实的理论和实验基础。在国家自然科学基金的资助下，项目进展顺利并取得了较好的研究成果。我们成功地研制出了符合实验要求的低重频，纳秒脉冲的绿光光源，利用此光源，实现了荧光染料和激光输出。同时我们还制备了荧光蛋白表达的细胞系，通过尝试各种培养方法，使细胞能够更好的发光。在此基金资助下，我们还建立了微结构光纤的生物传感平台，可以灵敏地检测各种荧光标记物。利用纳米材料的光学性能，比如光热转换和光敏毒性，我们还研究了肿瘤的早期检测和治疗，都取得了很好的成果。在此基金项目资助下，共获得研究成果33项。培养硕士生11名，其中5名已经取得硕士学位，6名在读。项目投入经费81万元，支出64.95万元(另有一实验材料5600元已经购买但是未报销，以及一项专利代理费3500已经办理借款，此两笔支出均发生在2017年12月31日之前)，各项支出基本与预算相符。剩余经费16.05万元，剩余经费计划用于本项目研究后续支出。