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›› 2016, Vol. 38 ›› Issue (3): 236-243.

• 论文与报告 • 上一篇    下一篇

一种快捷的单细胞藻类计数方法——运用Photoshop软件分析细胞显微图像轮廓

李茉莉1,乔琨2   

  1. 1. 湖南农业大学
    2. 福建省水产研究所
  • 收稿日期:2016-03-31 修回日期:2016-04-21 出版日期:2016-06-27 发布日期:2016-07-18
  • 通讯作者: 乔琨
  • 基金资助:
    海洋经济创新发展区域示范项目;福建省海洋经济创新发展区域示范项目;厦门南方海洋研究中心项目

A method of quick determination of unicellular algae counting——using Photoshop software to analyze microscopic image contour model

1,   

  • Received:2016-03-31 Revised:2016-04-21 Online:2016-06-27 Published:2016-07-18

摘要: 细胞计数是藻类培养、赤潮调查和环境监测中的一个重要环节。本文设计并测试了一种单细胞藻类计数方法:利用体视显微镜及配套的CCD数字成像系统,获取固定体积样品的全视野图像,运用Photoshop软件对细胞显微图像轮廓进行分析计数。根据细胞粒径差异,实验选取5种代表性单细胞藻类,分别采用新方法和显微镜计数法对其进行细胞密度测定。研究结果表明,新方法在测定亚心形扁藻(Platymonas subcordiformis)时,结果较显微镜计数明显偏低。而在测定东海原甲藻(Prorocentrum donghaiense)、米氏凯伦藻(Karenia mikimotoi)、塔玛亚历山大藻(Alexandrium tamarense)和血红哈卡藻(Akashiwo sanguinea)时,计数结果与显微镜计数结果非常接近,较可信。其计数结果与显微镜计数结果具有较好的线性相关性(R2≥0.981 1,P<0.01)。新方法能够有效测定粒径15 μm以上纯种或混合培养的单细胞藻类的生物量,简化了测定程序的同时,减少了人为误差,具有一定的应用价值。

关键词: 单细胞藻类, 藻类计数方法, Photoshop软件分析, 细胞显微图像轮廓, unicellular algae, cell counting method, Photoshop software analysis, microscopic image contour model

Abstract: Cell counting is an important step in algae culture, red tide investigation and water environment monitoring. A quick counting method of single cell algae was proposed and tested: the image of fixed volume sample was captured by the stereomicroscope and imaging instrument. And then Photoshop software was used to analyze the contour model of microscopic image. In this study, 5 representative microalgae species with different cell sizes were selected and the cell densities were measured by our new method and conventional microscope counting method, respectively. The results indicated that the counted numbers of cell density by our new method were apparently lower than those by microscope measurement for Platymonas subcordiformis. However, for other four algae species (Prorocentrum donghaiense, Karenia mikimotoi, Alexandrium tamarense, Akashiwo sanguinea), the counting results of the two methods were quite close, presenting a significant linear correlation between them (R2≥0.981 1). Thus, for unicellular alga with sizes larger than 15 μm, the new method could determine the biomass of pure and mixed cultivation effectively. Compared with the microscope counting method, the new method simplifies the measure procedures and reduces the artificial errors. Therefore, it has a certain application value in counting of single-cell or particles.

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