skip to primary navigationskip to content
 

A concept for single-shot volumetric fluorescence imaging via orthogonally polarized excitation lattices

Ströhl F,  Kaminski CF. "A concept for single-shot volumetric fluorescence imaging via orthogonally polarized excitation lattices."Sci. Rep. (2019) 9(1):6425. DOI |pdf


Abstract

The deconvolution of widefeld fuorescence images provides only guesses of spatial frequency information along the optical axis due to the so called missing cone in the optical transfer function. Retaining the single-shot imaging speed of deconvolution microscopy while gaining access to missing cone information is thus highly desirable for microscopy of volumetric samples. Here, we present a concept that superimposes two orthogonally polarized excitation lattices with a phase-shift of p between them. In conjunction with a non-iterative image reconstruction algorithm this permits the restoration of missing cone information. We show how fuorescence anisotropy could be used as a method to encode and decode the patterns simultaneously and develop a rigorous theoretical framework for the method. Through in-silico experiments and imaging of fxed biological cells on a structured illumination microscope that emulates the proposed setup we validate the feasibility of the method.