Fluorescence microscopy is the gold standard for investigation of biological samples. While powerful, this technology has its limitations, including phototoxicity, technical difficulties in introducing fluorescent markers, and limited simultaneous labeling of different structures due to the need for spectral separation. Here, artificial intelligence-driven virtual staining can provide label-free imaging. Deep neural networks are trained to learn the correlation between a label-free image and a ground-truth fluorescence image. However, the main challenge for the transition from traditional to virtual staining is the acquisition of huge datasets including labeled and unlabeled data. We have performed proof-of-concept experiments to investigate the possibilities of transfer learning in virtual staining. Therefore, U-Net architectures were pretrained on a larger dataset and later on trained again on a smaller and more specific dataset. We show that transfer learning decreases the needed dataset size and may even improve prediction quality. Furthermore, the interpretability of the trained networks was studied.This was investigated using guided backpropagation and modified input images.