PAExM: label-free hyper-resolution photoacoustic expansion microscopy.

Reflection-mode ultraviolet photoacoustic microscopy (UV-PAM) is capable of imaging cell nuclei in thick tissue without complex preparation procedures, but it is challenging to distinguish adjacent nuclei due to the limited spatial resolution. Tissue expansion technology has recently been developed to exceed the diffraction-limited fluorescence microscopies, but it is accompanied by limitations including additional staining. Herein, photoacoustic expansion microscopy (PAExM) is presented, which is an advanced histologic imaging strategy combining advantages of fast label-free reflection-mode UV-PAM and the tissue expansion technology. Clustered cell nuclei in an enlarged volume of a mouse brain section can be visually resolved without staining, demonstrating a great potential of the system to be widely used for histologic applications throughout biomedical fields.