The data augmentation method is known as a helpful technique to generate a dataset with a large number of images from one with a small number of images for supervised training in deep learning. However, a low validity augmentation method for image recognition was reported in a recent study on artificial intelligence (AI). This study aimed to clarify the optimal data augmentation method in deep learning model generation for the recognition of white blood cells (WBCs). Study Design: We conducted three different data augmentation methods (rotation, scaling, and distortion) on original WBC images, with each AI model for WBC recognition generated by supervised training. The subjects of the clinical assessment were 51 healthy persons. Thin-layer blood smears were prepared from peripheral blood and subjected to May-Grünwald-Giemsa staining. Results: The only significantly effective technique among the AI models for WBC recognition was data augmentation with rotation. By contrast, the effectiveness of both image distortion and image scaling was poor, and improved accuracy was limited to a specific WBC subcategory. Conclusion: Although data augmentation methods are often used for achieving high accuracy in AI generation with supervised training, we consider that it is necessary to select the optimal data augmentation method for medical AI generation based on the characteristics of medical images.