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利用流式细胞术比较九种甲藻的脂质含量

Comparison of Lipid Content in Nine Dinoflagellate Species Using Flow Cytometry.

作者信息

Park Jaeyeon, Yoon Eun Young, Moon Seung Joo, Hyung Jun-Ho, Lee Hangy

机构信息

Advanced Institute of Convergence Technology, Suwon 16229, Republic of Korea.

出版信息

Microorganisms. 2024 Dec 30;13(1):44. doi: 10.3390/microorganisms13010044.

Abstract

The lipid content of nine dinoflagellates was analyzed using flow cytometry to compare lipid levels. Additionally, the correlation between lipid content, cell size, and carbon content in dinoflagellates was evaluated using BODIPY 505/515 staining. The flow cytometry side scatter (SSC) effectively represented relative cell size, showing a linear relationship with the equivalent spherical diameter (ESD). Larger cells exhibited higher relative lipid content; however, lipid accumulation was influenced by nutritional modes and habitats, with mixorophic and benthic species displaying higher lipid content than heterotrophic species. A comparison of fluorescent dyes revealed that Nile Red overestimated lipid content, suggesting overlap with chlorophyll autofluorescence. Flow cytometry analysis with BODIPY 505/515 demonstrated a linear correlation with the sulfo-phospho-vanillin (SPV) method, enabling determination of actual lipid content using FL1 fluorescence and the slope value. As the carbon content increased, the lipid content initially increased rapidly but plateaued at higher carbon levels, indicating saturation. These findings suggest that relative fluorescence via flow cytometry provides an effective means to estimate the lipid content and carbon content as a function of cell size.

摘要

使用流式细胞术分析了九种甲藻的脂质含量,以比较脂质水平。此外,使用BODIPY 505/515染色评估了甲藻脂质含量、细胞大小和碳含量之间的相关性。流式细胞术侧向散射(SSC)有效地代表了相对细胞大小,与等效球径(ESD)呈线性关系。较大的细胞表现出较高的相对脂质含量;然而,脂质积累受营养模式和栖息地的影响,混合营养型和底栖物种的脂质含量高于异养物种。荧光染料比较显示,尼罗红高估了脂质含量,表明与叶绿素自发荧光存在重叠。使用BODIPY 505/515进行的流式细胞术分析表明与磺基磷酸香草醛(SPV)方法呈线性相关,从而能够使用FL1荧光和斜率值测定实际脂质含量。随着碳含量增加,脂质含量最初迅速增加,但在较高碳水平时趋于平稳,表明达到饱和。这些发现表明,通过流式细胞术的相对荧光提供了一种有效的方法来估计脂质含量以及作为细胞大小函数的碳含量。

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