| Summary: | Verifying pollen germination using microscopic images is a difficult task. It is usually time-consuming and may entail reduced accuracy and reproducibility. Therefore, in this study, we used random forest (RF) and convolutional neural network (CNN) models to perform image classification on raw data corresponding to pollens with different germination rates; the data were obtained via flow cytometry. A heat map, which was based on the RF analysis results, showed that the variables that significantly influenced the classification decision between NG and 60G categories were mainly located in the center and top-right regions of the <inline-formula> <tex-math notation="LaTeX">$30\times30$ </tex-math></inline-formula> pixel image. Additionally, a variable importance plot showed that among the 900 input variables, pixel_316 was the variable that contributed the most toward prediction. Gradient-weighted class activation mapping was used to visualize the class activation maps of the CNN model. The bottom-left region of the activation map was activated in the NG image. However, the 60G image showed that not only the bottom-left region but also the top-right region was activated. Both the models classified the input images into NG and 60G categories with high accuracy. However, considering that the RF model does not reflect the characteristics of adjacent variables, the CNN model is more appropriate for classifying pollen germination images corresponding to pollen with various germination rates into distinct classes. Taken together, these results suggest that the CNN model can provide a reliable method for verifying the pollen performance.
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