基于光流控芯片的船舶压载水活性微藻快速检测机理研究

With the gradual implementation of International Convention for the Control and Management of Ship's Ballast and Sediment, fast on-site detection of single microalgae cell activity has become one of the most important and burning questions. At present, there is still not a suitable method and device for detecting ballast water on-site. To fill in this gap, in this project, a fast on-site detection mechanism of single microalgae activity based on a novel principle is presented. The basic principle is the optical elements are integreted into the microfluodic channel using the advanced optofludic method and the single microalgae cells are manipulated and light induced chlorophyll delayed fluorescence (LCDF) is excited. Mechanism of LCDF origin and characterization of the photosynthesis activity of microalgae cell are studied and optimal LCDF parameters and a database of relationship between LCDF characteristics and cell activities are setup. This method has some advantages such as quickness, accuracy, label-free, simple operation, portable and on-site examination and so on. This method will have a wide application perspective. As research foundation, the applicant and his research group have published a number of related papers and patents. The applicant and his group have research characteristics and advantages in the multidisciplines of micro/optofluodic chip and ship ocean engineering.

随着《船舶压载水及沉积物控制和管理国际公约》的逐步实施，船舶压载水中活性微藻现场快速检测成为了急需解决的重要问题之一。为填补当前缺少适合船舶压载水现场快速检测方法与设备的空白，本课题提出一种新的压载水中活性微藻现场快速检测机理- - 利用国际微流控芯片领域先进的光流控方法,将光学元器件集成进入微通道内，通过光与微藻细胞的作用机制，在微通道内对单个微藻细胞进行操控，激发微藻体内光诱导叶绿素延迟荧光（LCDF），探究此荧光来源及表征微藻活性机理，分析LCDF荧光特性并筛选特征参数，建立LCDF荧光特性参数与微藻活性关联理论模型与数据库。该方法具有快速、准确、免标记、操作简单、便携、现场检测等优点，具有广泛的应用前景。作为研究基础，本课题申请人及其课题组已发表多篇相关期刊论文与专利，在微/光流控芯片及船舶海洋工程交叉领域具有研究特色与优势。

船舶压载水是外来海洋生物入侵的主要途径之一，外来有害海洋生物入侵传播造成的灾害，对生态、工业、农业以及人类健康等都造成了严重的影响。目前现存方法均无法从根本上解决船舶压载水单微藻活性现场快速检测的问题。在国家自然科学基金青年项目“基于光流控芯片的船舶压载水活性微藻快速检测机理研究”中，本课题组主要研究了在微流控芯片上通过激光激发微藻细胞使其产生光诱导叶绿素荧光（Light Induced Chlorophyll Fluorescence, LICF），探究了此荧光的来源及表征微藻活性机理，分析了LICF荧光特性并筛选特征参数，通过对LICF衰减动力学曲线分析，得出了衰减动力学曲线下积分可作为评价微藻细胞活性的参数，建立了LICF荧光特性参数与微藻活性关联理论模型，解决了诱导单微藻细胞LICF的参量阈值条件，单微藻细胞LICF激发与辐射方向性问题，以及单微藻细胞LICF与活性关联问题，实现了一种新的基于光流控芯片和LICF传感的船舶压载水中动态单微藻细胞活性快速检测方法并构建基于该方法的传感器，具有快速、准确、现场、便携等优点。本课题组在项目研究过程中取得的研究成果包括：发表相关论文13篇，其中SCI收录期刊论文10篇，EI收录论文3篇，申请国家发明专利9项，其中获得授权4项，在审5项，获得授权实用新型专利1项，培养了博士毕业研究生1名，硕士毕业研究生6名，在读博士研究生2名，在读硕士研究生4人。本课题组参与主办了2016年大连国际微纳流体和芯片实验室会议，并参加其他国际会议2人次。