V-shaped PSF for 3D imaging over an extended depth of field in wide-field microscopy.

Single-shot 3D optical microscopy that can capture high-resolution information over a large volume has broad applications in biology. Existing 3D imaging methods using point-spread-function (PSF) engineering often have limited depth of field (DOF) or require custom and often complex design of phase masks. We propose a new, to the best of our knowledge, PSF approach that is easy to implement and offers a large DOF. The PSF appears to be axially V-shaped, engineered by replacing the conventional tube lens with a pair of axicon lenses behind the objective lens of a wide-field microscope. The 3D information can be reconstructed from a single-shot image using a deep neural network. Simulations in a 10× magnification wide-field microscope show the V-shaped PSF offers excellent 3D resolution (<2.5 µm lateral and ∼15 µm axial) over a ∼350 µm DOF at a 550 nm wavelength. Compared to other popular PSFs designed for 3D imaging, the V-shaped PSF is simple to deploy and provides high 3D reconstruction quality over an extended DOF.