ABSTRACT
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.