Simultaneous bright field and fluorescence dual optical path imaging of phytoplankton

XU Min; YIN Gaofang; ZHAO Nanjing; HU Xiang; HUANG Peng

doi:10.37188/OPE.20233112.1725

您当前的位置：

首页 >

文章列表页 >

Simultaneous bright field and fluorescence dual optical path imaging of phytoplankton

Modern Applied Optics | 更新时间：2026-09-11

- Simultaneous bright field and fluorescence dual optical path imaging of phytoplankton
- Optics and Precision Engineering Vol. 31, Issue 12, Pages: 1725-1734(2023)
- 作者机构：
  
  1.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
  2.中国科学技术大学，安徽合肥 230026
  3.合肥学院，安徽合肥 230601
- 作者简介：
- 基金信息：
- DOI：10.37188/OPE.20233112.1725
  CLC： X832
- Received：03 November 2022，
  
  Revised：2022-12-07，
  
  Published：25 June 2023
- 稿件说明：
移动端阅览
徐敏,殷高方,赵南京等.浮游植物明场与荧光双光路同步成像方法[J].光学精密工程,2023,31(12):1725-1734.

XU Min,YIN Gaofang,ZHAO Nanjing,et al.Simultaneous bright field and fluorescence dual optical path imaging of phytoplankton[J].Optics and Precision Engineering,2023,31(12):1725-1734.
徐敏,殷高方,赵南京等.浮游植物明场与荧光双光路同步成像方法[J].光学精密工程,2023,31(12):1725-1734. DOI： 10.37188/OPE.20233112.1725.

XU Min,YIN Gaofang,ZHAO Nanjing,et al.Simultaneous bright field and fluorescence dual optical path imaging of phytoplankton[J].Optics and Precision Engineering,2023,31(12):1725-1734. DOI： 10.37188/OPE.20233112.1725.

摘要

针对目前浮游植物种类鉴别镜检法中存在的相似形态藻细胞难以区分、同粒径杂质干扰大、藻细胞位置难以定位分割等问题，设计了一种浮游植物明场荧光显微同步成像系统。系统使用高分辨显微成像光路结构、透射式与落射式结合的照明方式和以分光光路为核心的明场荧光双光路同步成像方法，获取浮游植物显微图像下的明场形态信息和色素荧光信息。利用该系统对绿藻门、蓝藻门、硅藻门等不同门类藻种进行明场图像与荧光图像同步成像测试，实验结果表明：系统成像分辨率达1.5 μm，成像周期时间为707 ms，系统采集的图片中藻细胞纹理清晰，细胞边缘明显，利用系统中的明场图像并结合荧光图像，可有效定位藻细胞位置，降低样品中杂质对藻种鉴别的影响，丰富浮游植物显微图像信息量，为基于浮游植物显微图像的相关研究提供方法参考。

Abstract

A phytoplankton bright-field fluorescence microscopic simultaneous imaging system was designed to solve the problems of the conventional phytoplankton species identification microscopy method， such as the difficulty of distinguishing between similar morphological algal cells， interference of impurities of the same particle size， and difficulty in locating and segmenting the algal cells. The proposed system uses a high-resolution microscopic imaging optical path structure， combined transmission-fall illumination method， and simultaneous bright-field fluorescence dual-optical imaging method with a spectral optical path to capture bright-field morphological and fluorescence images of phytoplankton with pigment information. The system was tested for the simultaneous acquisition of bright-field and fluorescence images of different algal species， including chlorophyta， cyanophyta， and bacillariophyta. The experimental results indicated that the imaging resolution of the system was 1.5 μm， imaging period was 707 ms， texture of algal cells in the images captured by the system was clear， and cell edges were obvious. The system using the method of combining the bright-field images with fluorescent images can effectively locate the algal cells， reduce the influence of impurities in the samples on the procedure of algae identification， and increase the amount of information obtained from the algal microscopic images. This study provides a methodological reference for related research based on phytoplankton microscopic images.

关键词

Keywords

references

TANG D L ， DI B P ， WEI G F ， et al . Spatial， seasonal and species variations of harmful algal blooms in the South Yellow Sea and East China Sea ［J］. Hydrobiologia ， 2006 ， 568 （ 1 ）： 245 - 253 . doi: 10.1007/s10750-006-0108-1 http://dx.doi.org/10.1007/s10750-006-0108-1

KHAMMERI Y ， BELLAAJ-ZOUARI A ， HAMZA A ， et al . Ultraphytoplankton community composition in Southwestern and Eastern Mediterranean Basin： relationships to water mass properties and nutrients ［J］. Journal of Sea Research ， 2020 ， 158 ： 101875 . doi: 10.1016/j.seares.2020.101875 http://dx.doi.org/10.1016/j.seares.2020.101875

YANG S M ， LIU X S . Characteristics of phytoplankton assemblages in the southern Yellow Sea， China ［J］. Marine Pollution Bulletin ， 2018 ， 135 ： 562 - 568 . doi: 10.1016/j.marpolbul.2018.07.062 http://dx.doi.org/10.1016/j.marpolbul.2018.07.062

EİDEMIR ， TURKOGLU M . Temporal variations of phytoplankton community and their correlation with environmental factors in the coastal waters of the Çanakkale Strait in 2018 ［J］. Oceanologia ， 2022 ， 64 （ 1 ）： 176 - 197 . doi: 10.1016/j.oceano.2021.10.003 http://dx.doi.org/10.1016/j.oceano.2021.10.003

BEDFORD J ， OSTLE C ， JOHNS DG ， et al . The influence of temporal scale selection on pelagic habitat biodiversity indicators ［J］. Ecological Indicators ， 2020 ， 114 ： 106311 . doi: 10.1016/j.ecolind.2020.106311 http://dx.doi.org/10.1016/j.ecolind.2020.106311

张军毅，孙蓓丽，朱冰川，等 . 基于分子标记的藻类鉴定研究进展［J］. 湖泊科学， 2021 ， 33 （ 6 ）： 1607 - 1625 . doi: 10.18307/2021.0601 http://dx.doi.org/10.18307/2021.0601

ZHANG J Y ， SUN B L ， ZHU B C ， et al . Research progress in the taxonomic identification of algae on the basis of molecular markers ［J］. Journal of Lake Sciences ， 2021 ， 33 （ 6 ）： 1607 - 1625 . （in Chinese） . doi: 10.18307/2021.0601 http://dx.doi.org/10.18307/2021.0601

COLTELLI P ， BARSANTI L ， EVANGELISTA V ， et al . Water monitoring： automated and real time identification and classification of algae using digital microscopy ［J］. Environmental Science Processes & Impacts ， 2014 ， 16 （ 11 ）： 2656 - 2665 . doi: 10.1039/c4em00451e http://dx.doi.org/10.1039/c4em00451e

WALKER R F ， ISHIKAWA K ， KUMAGAI M . Fluorescence-assisted image analysis of freshwater microalgae ［J］. Journal of Microbiological Methods ， 2002 ， 51 （ 2 ）： 149 - 162 . doi: 10.1016/s0167-7012(02)00057-x http://dx.doi.org/10.1016/s0167-7012(02)00057-x

HENSE B A ， GAIS P ， JÜTTING U ， et al . Use of fluorescence information for automated phytoplankton investigation by image analysis ［J］. Journal of Plankton Research ， 2008 ， 30 （ 5 ）： 587 - 606 . doi: 10.1093/plankt/fbn024 http://dx.doi.org/10.1093/plankt/fbn024

DEGLINT J L ， JIN C ， CHAO A ， et al . The feasibility of automated identification of six algae types using feed-forward neural networks and fluorescence-based spectral-morphological features ［J］. IEEE Access ， 2018 ， 7 ： 7041 - 7053 . doi: 10.1109/access.2018.2889017 http://dx.doi.org/10.1109/access.2018.2889017

BI X ， LIN S ， ZHU S ， et al . Species identification and survival competition analysis of microalgae via hyperspectral microscopic images ［J］. Optik ， 2019 ， 176 ： 191 - 197 . doi: 10.1016/j.ijleo.2018.09.077 http://dx.doi.org/10.1016/j.ijleo.2018.09.077

LAUFFER M ， GENTY F ， COLLETTE J L ， et al . Phytoplankton Identification by Combined Methods of Morphological Processing and Fluorescence Imaging ［C］. 2015 IEEE Workshop on Environmental， Energy， and Structural Monitoring Systems （EESMS） Proceedings . 9 - 10 ， 2015， Trento， Italy. IEEE ， 2015： 131 - 135 . doi: 10.1109/eesms.2015.7175865 http://dx.doi.org/10.1109/eesms.2015.7175865

张朋涛，杨西斌，周伟，等 . 双模切换显微内窥镜成像系统设计及应用［J］. 光学精密工程， 2019 ， 27 （ 6 ）： 1335 - 1344 . doi: 10.3788/ope.20192706.1335 http://dx.doi.org/10.3788/ope.20192706.1335

ZHANG P T ， YANG X B ， ZHOU W ， et al . Design and applied research of dual-mode switching endomicroscopic imaging system ［J］. Opt. Precision Eng. ， 2019 ， 27 （ 6 ）： 1335 - 1344 . （in Chinese） . doi: 10.3788/ope.20192706.1335 http://dx.doi.org/10.3788/ope.20192706.1335

王雪，刘虹遥，路鑫超，等 . 无透镜全息显微细胞成像［J］. 光学精密工程， 2020 ， 28 （ 8 ）： 1644 - 1650 .

WANG X ， LIU H Y ， LU X C ， et al . Cell imaging by holographic lens-free microscopy ［J］. Opt. Precision Eng. ， 2020 ， 28 （ 8 ）： 1644 - 1650 . （in Chinese）

LU Q ， LIU G H ， XIAO C L ， et al . A modular， open-source， slide-scanning microscope for diagnostic applications in resource-constrained settings ［J］. PLoS One ， 2018 ， 13 （ 3 ）： e0194063 . doi: 10.1371/journal.pone.0194063 http://dx.doi.org/10.1371/journal.pone.0194063

缪新，张运海，黄维 . 皮肤反射式共聚焦显微成像自适应图像亮度调节［J］. 光学精密工程， 2019 ， 27 （ 6 ）： 1270 - 1276 . doi: 10.3788/ope.20192706.1270 http://dx.doi.org/10.3788/ope.20192706.1270

MIAO X ， ZHANG Y H ， HUANG W . Image brightness adaptive adjustment during skin imaging by reflectance confocal microscopy ［J］. Opt. Precision Eng. ， 2019 ， 27 （ 6 ）： 1270 - 1276 . （in Chinese） . doi: 10.3788/ope.20192706.1270 http://dx.doi.org/10.3788/ope.20192706.1270

殷高方，赵南京，胡丽，等 . 基于色素特征荧光光谱的浮游植物分类测量方法［J］. 光学学报， 2014 ， 34 （ 9 ）： 0930005 .

YIN G F ， ZHAO N J ， HU L ， et al . Classification and measurement method of phytoplankton based on pigment characteristic fluorescence spectrum ［J］. Acta Optica Sinica ， 2014 ， 34 （ 9 ）： 0930005 . （in Chinese）

贾仁庆，殷高方，赵南京，等 . 浮游藻类细胞显微明场图像与荧光同步测量图像配准方法研究［J］. 中国激光， 2022 ， 49 （ 24 ）： 80 - 87 .

JIA R Q ， YIN G F ， ZHAO N J ， et al . Registration method of microscopic bright field and fluorescence synchronous measurement images of phytoplankton cells ［J］. Chinese Journal of Lasers ， 2022 ， 49 （ 24 ）： 80 - 87 . （in Chinese）

Views

836

下载量

CSCD

Alert me when the article has been cited

提交

Tools

Publicity Resources

OPE cover | Phytoplankton bright field and fluorescence dual optical path synchronous imaging method

Birefringence microscopy imaging based on photoelastic modulation with stroboscopic illumination

Detection and validation of plant chlorophyll fluorescence employing Scheimpflug lidar

Design of gene sequencing fluorescence microscopic system with dual-band LED illumination

Daul-optical path observation system for tracking free-moving C. elegans

Autofocus of microarray digital PCR fluorescent chip

Related Author

LI Kewu

WANG Shuang

WANG Zhibin

REN Shuxuan

DONG Ye

MEI Liang

YANG Shuqing

ZHENG Kexin

Related Institution

Shanxi Provincial Engineering Technology Research Center for Optoelectronic Information and Instruments

College of Computer Science and Technology， North University of China

School of Electrical and Control Engineering， North University of China

Institute of Instrumentation and Electronics， North University of China

School of Optoelectronic Engineering and Instrumentation Science， Dalian University of Technology

AI问答

Address：No.3888 Dong Nanhu Road, Changchun, Jilin, China Postal code：130033
Tel：0431-86176855 Email：gxjmgc@ciomp.ac.cn
Technical support is provided by Beijing Founder electronics co., LTD 吉ICP备11002662号-17 京公网安备11010802024621
It is recommended to read the content of this site in Chrome&IE9+. Please switch to extreme mode in browser 360.
Cookies We use cookies to help provide and enhance our service and tailor content. By continuing, you agree to the use of cookies.

⁰