基于可变荧光统计分析法的混合藻类活体细胞数定量实验研究

Quantitatively Experimental Study on Number of Viable Cells in Mixed Algae Based on Variable Fluorescence Statistical Analysis

以混合藻类为研究对象,研究组成混合样品藻种的单细胞可变荧光量(SCVF)差异对可变荧光统计分析法准确性的影响。研究结果表明:当两种藻种的SCVF比值不大于5.3时,该分析方法依旧适用,测量结果与显微镜检测结果的相对误差绝对值的均值不大于17%;当两种藻种的SCVF比值大于8时,测量结果与显微镜检测结果存在较大差异,且高SCVF藻细胞占比达到一定比例时,可变荧光统计分析法的测量结果仅反映了混合样品中高SCVF藻细胞数。针对SCVF差异大的混合藻类,将过滤分离方法与可变荧光统计分析法相结合,利用合适孔径的金属滤网过滤混合样品,将过滤前后测量的样品活体藻细胞密度之和作为测量结果。实验结果表明,该方法可将相对误差绝对值均值由58.4%降至5.5%,有效解决了组成混合样品藻种的SCVF差异对细胞数定量的影响,实现混合藻类活体细胞数的准确定量。

This paper takes mixed algae as the research object to study the influence of single cell variable fluorescence(SCVF)differences of algal species in mixed samples on the accuracy of variable fluorescence statistical analysis.The research results show that when the SCVF ratio of two algae is no more than 5.3,the analysis is still applicable,and the mean absolute value of the relative errors between measurement results and microscopic examination results is no more than 17%.When the SCVF ratio of two algae is higher than 8,the measurement results are quite different from the microscopic examination results.In addition,when high SCVF algal cells account for a large proportion,the measurement results of variable fluorescence statistical analysis only reflect the number of high SCVF algal cells in the mixed samples.As a result,for the mixed algae with large differences in SCVF,the paper combines a filtration separation method with the variable fluorescence statistical analysis method.The mixed samples are filtered by a metal filter screen with suitable pore size,and the density sum of measured viable algal cells before and after filtration is taken as experimental result.The results show that the proposed method can greatly reduce the mean absolute value of the relative error from 58.4%to 5.5%,effectively mitigate the influence of the SCVF difference of algal species in mixed samples on the quantitative analysis of cell number,and accurately quantify viable cells in mixed algae.