Based on the sclereids in the bark of oak species, a convolutional neural network (CNN) was employed to validate species classification performance and its influencing factors. Three optimizers including stochastic gradient descent (SGD), adaptive moment estimation (Adam), root mean square propagation (RMSProp), and dataset augmentation were adopted. The accuracy and loss stabilized at approximately 15 to 20 and 70 to 80 epochs for the augmented and non-augmented condition, respectively. In the last five epochs, the RMSProp-augmented condition achieved the highest accuracy of 89.8%, whereas the Adam-augmented condition achieved the lowest accuracy of 73.8%. Regarding the loss, SGD-non-augmented condition was the lowest at 0.498, whereas Adam-augmented condition was the highest at 2.740. The highest accuracy was influenced by RMSProp at 0.194. Dataset augmentation had a significant influence on accuracy at 0.456. Homogeneous subsets among the validation conditions indicated that the accuracy and loss were classified into the same subset using an augmented dataset during the training, regardless of the optimizer. Only Adam and RMSProp with non-augmented datasets were categorized into the same subset during the test. Hence, species classification using CNN and sclereid characteristics in the bark was feasible, and RMSProp with augmented datasets showed optimal performance for species classification.