海洋微藻研究对海洋环境监测以及生物资源利用有着重要意义, 显微共焦拉曼光谱作为一种非标记、 快速检测技术已在生命领域获得广泛应用。现阶段, 商业化显微共焦拉曼仪器在微生物研究领域占据主导, 但由于体积庞大、工作环境要求严格等因素, 很难开展微藻细胞的现场检测与分析。因此, 将微型 光纤光谱仪引入微藻“单细胞”的检测, 自主研发了一套小型显微共焦拉曼系统, 尝试低成本开发微 生物分析设备。整个系统基于微型光纤光谱仪实现了硬件的一体化小型设计(L750 mm×W350 mm×H410 mm), 具备光谱探测、显微成像、光镊捕获功能。通过四种典型微藻(中肋骨条藻、微拟球藻、东海原甲藻及小藻)的检测验证, 成功识别了“单个活体细胞”内的蛋白质、脂类、糖原、核酸等多种细胞组分, 相应的结果经过主成分分析 (Principal component analysis, PCA)后, 很好地实现了四种微藻的种类归属, 证明了光纤光谱仪应用于单细胞量级 海洋微生物分析的可行性, 并有望在将来发展成为船基设备, 用于微藻的甲板在线检测。

The study of algae is significant for monitoring marine environment and exploring biological resources. Confocal Raman spectroscopy has been widely used in biology as a label-free and rapid detection technique. Currently, the commercial micro-Raman is dominated in this field, while it is not applicable to field measurements for the instrumental size and the operation condition. Hence, a fiber optic spectrometer is introduced to develop a low-cost compact confocal-raman system for algae detection at “single-cell” level. The developed system is all-in-one designed for the compactness, and is well functioned with the capacities of Raman detection, microscopic imaging and optical tweezers capture. Four typical kinds of single-cells (Skeletonema costatum, N.oceanica IMET-1, Prorocentrum donghaiense Lu and Chlorella) were used to evaluate the system detectability, and the components such as proteins, lipids, glycogen and nucleic acids were recognized. Based on that, the algae cells were successfully classified into 4 species after using principal component analysis (PCA) method, which indicates that it is feasible to apply the fiber optic spectrometer for single cell analysis of marine microbes. It is expected that the developed system will be deployed as field instrument for the deck measurements in sea trials in the future.