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目的：探讨基于单粒子倍性检测技术中光斑与细胞的相对大小对细胞倍性分析结果的影响，确定合适的入射场光斑范围。方法首先，对影响单粒子细胞倍性检测结果的主要因素光斑厚度进行理论分析；然后借助ZEMAX进行光束整形系统设计，通过仿真以优化整形系统的参数；最后根据细胞粒径，确立目标光斑尺寸，搭建光学检测平台，获取一组厚度不同的光斑，分别进行细胞倍性检测。结果仿真和光学平台均获得了目标光斑，细胞倍性实验中，光斑厚度同时大于单核、双联体细胞时，平均荧光强度比值为2．03，符合倍性指标要求。结论以电压脉冲信号峰高表征荧光强度时，光斑与细胞的相对大小会影响倍性检测的准确性，光斑厚度必须同时大于单倍体与多倍体细胞，才能得到真实的倍性关系。“,”Objective To explore the relationship between the spot size and the result of ploidy analysis in the detection technology based on single-particles, and to fix the scope of spot thickness on the basis of the experimental result.Methods The influence of spot thickness on voltage signals produced by single cells was analyzed.The parameters of the beam shap system in the incident field were designed and optimized on ZEMAX.Finally, according to the cells′diameter, the target size of spots was set.A set of spots of different thickness and of Gaussian distribution obtained from the optical experimental platform was used to conduct ploidy detection experiments.Results Target spots were both obtained from ZEMAX simulation and the optical platform.When the spot thickness was larger than both monocytes and coenocytes, the mean fluorescence intensity ratio was 2.03,which met the demand of the index.Conclusion When the height of the pulse is used to represent the fluorescence density, the relative size of spots and cells will affect the result of ploidy detection.Only when spot thickness is larger than cells is the ploidy ratio accurate.