文蛤和菲律宾蛤仔血细胞的图像流式分类分析  

Hemocytes Classification of Meretrix meretrix and Ruditapes philippinarum by Imaging Flow Cytometer

作　　者：任星潮 李荣 张宁 刘逸尘[1] 孙金生[1] 张亦陈[1] REN Xing-chao;LI Rong;ZHANG Ning(Tianjin Key Laboratory of Animal and Plant Resistance/College of Life Sciences,Tianjin Normal University,Tianjin 300387)

机构地区：[1]天津市动植物抗性重点实验室/天津师范大学生命科学学院,天津300387

出　　处：《安徽农业科学》2020年第8期98-101,共4页Journal of Anhui Agricultural Sciences

基　　金：天津市自然科学基金项目(19JCYBJC29700);天津市水产生态及养殖重点实验室开放基金项目(TJAE2015005);天津市高等学校创新团队建设规划项目(TD13-5076);天津市水产产业技术体系创新团队项目(ITTFRS2017007);天津师范大学高研高访项目。

摘　　要：[目的]探讨新型图像流式细胞仪在贝类血细胞分类中的应用。[方法]建立了一种基于血细胞侧向散射强度和面积测量的自动化方法,并与显微观察对比其在文蛤和菲律宾蛤仔血细胞分类中的效果。[结果]参考显微镜下可辨识的胞内颗粒结构,2种贝类的血细胞均可分为无颗粒、小颗粒及大颗粒3类,但基于人工辨识的统计结果缺乏量化标准,因此波动较大。图像流式细胞仪通过定量分析可以获得更精细、准确的测量结果。根据侧向散射强度和面积差异,将文蛤和菲律宾蛤仔的血细胞分成无颗粒、小颗粒、中颗粒及大颗粒4类,占比分别为(6.59±1.11)%和(30.03±1.87)%、(34.85±2.95)%和(58.50±5.73)%、(21.95±0.75)%和(6.03±1.44)%、(36.75±4.75)%和(5.36±2.38)%。2种贝类血细胞都呈现出由无颗粒细胞到大颗粒细胞逐渐增大的趋势。[结论]2种方法的分析结果既有联系又不完全一致。相较于显微观察,图像流式细胞仪利用精确的量化标准可辨识出更多的细胞类群,提示综合利用不同的特征差异还可以获得更精细的分类结果。此外,自动化的定量测量准确性好、通量高,是今后贝类血细胞分析的重要方法。[Objective]To discuss the application of a new kind of imaging flow cytometer in the classificatioin of shellfish hemocyte.[Method]An automated method based on the measurement of blood cell lateral scattering intensity and area was established,and its effect on hemocyte classification of Meretrix meretrix and Ruditapes philippinarum was compared with microscopic observation.[Result]Both of the hemocytes from the above shellfishes could be divided into 3 hemocyte populations of non-granular,small-granular and large-granular hemocyte by microscopic observation according to the differences of distinguishable intracellular particles,but the proportions fluctuated due to lacking of quantitative criteria.More accurate results could be acquired by using a quantitative measurement of imaging flow cytometer.According to the difference of the side scattering intensity and area,the hemocytes of M.meretrix and R.philippinarum were divided into four types:non-granular,small-granular,intermediate-granular and large-granular hemocytes,and their populations in M.meretrix and R.philippinarum were(6.59±1.11)%and(30.03±1.87)%,(34.85±2.95)%and(58.50±5.73)%,(21.95±0.75)%and(6.03±1.44)%,(36.75±4.75)%and(5.36±2.38)%respectively.The cell size increased gradually from R1 to R4 for each shellfish.[Conclusion]Although there are differences between the consistencies between two methods still could be found.Compared with microscopic observation,the image flow cytometer can identify more cell groups by using precise quantification standards,suggesting that more refined classification results could be obtained by using comprehensive use of different feature differences.In addition,automated quantitative measurement had good accuracy and high throughput,it is an important analysis method for blood cells of shellfish in the future.

关 键 词：文蛤 菲律宾蛤仔 血细胞 分类 流式细胞仪 

分 类 号：S917.4[农业科学—水产科学]