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

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Simultaneous bright field and fluorescence dual optical path imaging of phytoplankton

Modern Applied Optics | 更新时间：2023-07-17

- 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：07 December 2022，
  
  Published：25 June 2023
- 稿件说明：
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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.

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references

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