Multi-plane, wide-field fluorescent microscopy for biodynamic imaging in vivo

Wide-field fluorescent microscopy (WFFM) is widely employed in biomedical studies, due to its inherent advantages in high-speed imaging of biological dynamics noninvasively and specifically. However, WFFM suffers from the loss of axial resolution and the poor resistance to light scattering in deep tissue imaging. Here we propose a novel WFFM which has the capability in optical sectioning and volumetric imaging. We perform speckle illumination with a digital-micromirror-device for optical sectioning and employ an electrically tunable lens for defocusing modulation so as to quickly switch the image planes. We demonstrate its applications in multi-plane, wide-field imaging of biological dynamics in both zebrafish brains and mouse brains in vivo.