基于微流控-显微荧光技术的浮游藻细胞密度检测方法  

Microfluidics-Microfluorescence-Based Method for Density Detection of Planktonic Algal Cells

作　　者：黄朋 殷高方[1,2,3] 赵南京 甘婷婷 胡翔[1,2] 徐敏 梁天泓 贾仁庆 张小玲[2,4] Huang Peng;Yin Gaofang;Zhao Nanjing;Gan Tingting;Hu Xiang;Xu Min;Liang Tianhong;Jia Renqing;Zhang Xiaoling(School of Biological Food and Environment,Hefei University,Hefei 230601,Anhui,China;Key Laboratory of Environmental Optics and Technology,Anhui Institute of Optics and Fine Mechanics,Hefei Institutes of Physical Science,Chinese Academy of Sciences,Hefei 230031,Anhui,China;University of Science and Technology of China,Hefei 230026,Anhui,China;Institutes of Material Science and Information Technology,Anhui University,Hefei 23060l,Anhui,China)

机构地区：[1]合肥学院生物食品与环境学院,安徽合肥230601 [2]中国科学院合肥物质科学研究院安徽光学精密机械研究所中国科学院环境光学与技术重点实验室,安徽合肥230031 [3]中国科学技术大学,安徽合肥230026 [4]安徽大学物质科学与信息技术研究院,安徽合肥230601

出　　处：《光学学报》2023年第18期286-293,共8页Acta Optica Sinica

基　　金：国家重点研发计划项目(2021YFC3200100);安徽省科技重大专项(202203a07020002,202003a07020007);国家自然科学基金项目(62005001,61875207)。

摘　　要：浮游藻类密度监测对水质状况诊断及藻华灾害预警具有重要意义。因此,提出一种基于微流控-显微荧光技术的浮游藻细胞密度检测方法。该方法基于微流控技术实现样品快速定量进样,利用共聚焦显微荧光结构实现藻细胞特征荧光信号的高信噪比采集,并通过分析荧光峰信息实现浮游藻细胞计数。以杜氏盐藻、色球藻、隐藻和赤潮藻为测试对象的结果表明:在1.3×10^(6)L^(-1)密度范围内测量相对误差均小于3.96%,且准确率不受悬浮物、藻细胞种类以及尺寸的影响;在10%允许误差下,藻类密度检测上限可提升至5×10^(6)L^(-1),完全能够满足自然水体浮游藻细胞密度检测需求,为水体藻细胞密度快速准确检测提供了新途径。Objective As indicators of the ecological health of water bodies,planktonic algae are important primary producers in water ecosystems.The density monitoring of planktonic algae is of great significance to the diagnosis of water quality and the warning of algal blooms.Due to the presence of small individual and large numbers of planktonic algae,suspended impurities and other factors,traditional methods are difficult to achieve rapid and accurate measurements.Flow cytometric fluorescence method counts by detecting single cell fluorescence.This method features rapid,accurate,and highly efficient measurement,but it is not suitable for miniaturized field rapid measurement because of its complex injection structure and cumbersome focusing mode.Microfluidic chip technology realizes the functions of feeding,focusing,and sorting by constructing micro-channel pipelines on a square centimeter chip.This technology can simplify the complex feeding structure of the flow fluorescence method and has been widely employed in pharmaceutical and life science fields.Based on chlorophyll fluorescence in the characteristic band emitted by excited algal cells,this paper combines microfluidic chip technology and microfluorescence detection technology.It aims to realize rapid and accurate density detection of planktonic algal cells by detecting the number of single algal cell fluorescence peaks at a specific volume under a simple structure.Methods The experimental system consists of a sample feeding module,a fluorescence excitation module,and a fluorescence detection module.The excitation light from the monochromatic high-brightness LD is focused on the surface of the microfluidic channel by the drop-in microscopic optical structure.The algal cells in the microfluidic channel pass through the excitation window at a uniform speed under the propulsion of the syringe pump,and the cells are excited to emit fluorescence.Each cell flow across the microscopic field of view corresponds to a fluorescence peak,and then the density of algal cells in

关 键 词：浮游藻类 显微荧光 微流控 藻细胞计数 

分 类 号：X835[环境科学与工程—环境工程]