面向微藻单细胞检测的小型共焦拉曼系统研制

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。

Development of Compact Confocal Raman System for Algae Single-Cell Detection

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 detectiontechnique. Currently, the commercial micro-Raman is dominated in this field, while it is notapplicable 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 systemfor algae detection at ``single-cell' level. The developed system is all-in-one designed forthe compactness, and is well functioned with the capacities of Raman detection, microscopicimaging and optical tweezers capture. Four typical kinds of single-cells (Skeletonema costatum,N.oceanica IMET-1, Prorocentrum donghaiense LuandChlorella) were used to evaluate the systemdetectability, and the components such as proteins, lipids, glycogen and nucleic acids wererecognized. Based on that, the algae cells were successfully classified into 4 speciesafter using principal component analysis (PCA) method, which indicates that it isfeasible to apply the fiber optic spectrometer for single cell analysis of marinemicrobes. It is expected that the developed system will be deployed as field instrumentfor the deck measurements in sea trials in the future.