CAJ | 학술논문

微型浮游生物(细胞粒径<20μm)在水生生态系统的物质循环和能量流动中起着重要的作用，对其丰度的准确测定是进一步研究微型浮游生物在不同水生生态系统中作用的重要基础。相对于传统的显微镜检测技术，流式细胞术不仅具有分析速度快、灵敏度和准确度高等优点，而且可以同时测量单个细胞的多个生理参数。不同类型微型浮游生物流式细胞术的应用原理是不同的。对于自养型浮游藻类，主要根据藻体内色素的自发荧光对其进行分辨和计数；而对于异养型细菌、原生动物及浮游病毒等，还需借助外源荧光染料对细胞核酸染色后再进行分析。目前流式细胞术已成为浮游藻类和异养细菌丰度检测的常规方法，但是，由于原生动物具有更大的细胞体积且在自然水体中丰度较低；而浮游病毒粒径又太小，甚至低于光源激发波长，因此流式细胞术应用一直受到限制，直到近10年来才有相关报道。本文对运用流式细胞术计数浮游藻类、浮游细菌、原生动物和浮游病毒的具体原理、方法和进展进行综述，并对流式细胞仪在未来水生微生物学领域的应用进行展望。
미형부유생물(세포립경<20μm)재수생생태계통적물질순배화능량류동중기착중요적작용，대기봉도적준학측정시진일보연구미형부유생물재불동수생생태계통중작용적중요기출。상대우전통적현미경검측기술，류식세포술불부구유분석속도쾌、령민도화준학도고등우점，이차가이동시측량단개세포적다개생리삼수。불동류형미형부유생물류식세포술적응용원리시불동적。대우자양형부유조류，주요근거조체내색소적자발형광대기진행분변화계수；이대우이양형세균、원생동물급부유병독등，환수차조외원형광염료대세포핵산염색후재진행분석。목전류식세포술이성위부유조류화이양세균봉도검측적상규방법，단시，유우원생동물구유경대적세포체적차재자연수체중봉도교저；이부유병독립경우태소，심지저우광원격발파장，인차류식세포술응용일직수도한제，직도근10년래재유상관보도。본문대운용류식세포술계수부유조류、부유세균、원생동물화부유병독적구체원리、방법화진전진행종술，병대류식세포의재미래수생미생물학영역적응용진행전망。
The small plankton( cell size less than 20μm) plays an important role in the circulation of materials and energy flow of the aquatic system. Accurate enumeration of these organisms is the basis for understanding their ecological role in various water bodies. In comparison with microscopy observation, flow cytometry analysis is much more advantageous in terms of speed, sensitiv-ity and accuracy. Moreover, multiple parameters of a single cell could be measured simultaneously using flow cytometry. The proto-col to enumerate plankton depends on the type of plankton. Discrimination of several major groups of phytoplankton is mainly based on the differences in the fluorescence properties of their photosynthetic pigments. While for the heterotrophic bacteria, protozoan and viruses, a combination of exogenous fluorochromes staining on cell components( mainly nucleic acids) is required to better characterize different cell groups. Now flow cytometry has become a routine methodology for detecting density of the autotrophic phytoplankton and heterotrophic bacterioplankton. However, it has been only used in quantification of protozoan and viruses in the recent 10 years, for those applications which are much more difficult and complicated for the larger cell size and less abundant den-sities of protozoan and much smaller cell size( even smaller than the wavelength of the laser light used) of viruses compared to bac-terioplankton and small phytoplankton. The different principles and protocols used to discriminate autotrophic phytoplankton, het-erotrophic bacteria, protozoan and viruses through flow cytometry were reviewed in detail, and future applications of flow cytometry in aquatic microbiology were also prospected.