Imaging and positioning through scattering media with double-helix point spread function engineering

To date, super-resolution imaging through turbid media has not been reported.Aim: To explore the potential of DH-PSF microscopy in the imaging and localization of targets in scattering environments for improved 3D localization accuracy and imaging quality.Approach: The conventional DH-PSF method was modified to accommodate the scanning strategy combined with a deconvolution algorithm.

The localization of a fluorescent microsphere is determined by the center of the corresponding double spot, whereas the image is reconstructed from the scanned data by deconvoluting the DH-PSF.Results: The resolution, i.e. the localization accuracy, was calibrated to 13 nm in the transverse plane and 51 nm in the axial direction. Penetration thickness could reach an optical thickness (OT) of 5. Proof-of-concept imaging and the 3D localization of fluorescent microspheres through an eggshell membrane and an inner epidermal membrane of an onion were presented to demonstrate the super-resolution and optical sectioning capabilities.Conclusions: Modified DH-PSF microscopy can image and localize targets buried in scattering media using super-resolution.