Enhancing obSTORM imaging performance with cubic spline PSF modeling

Abstract

Oblique plane microscopy-based single molecule localization microscopy (obSTORM) has shown great potential for super-resolution imaging of thick biological specimens. Despite its compatibility with tissues and small animals, prior uses of the Gaussian point spread function (PSF) model have resulted in limited imaging resolution and a narrow axial localization range. This is due to the poor fit of the Gaussian PSF model with the actual PSF shapes in obSTORM. To overcome these limitations, we have employed cubic splines for a more accurate modeling of the experimental PSF shapes. This refined PSF model enhances three-dimensional localization precision, leading to significant improvements in obSTORM imaging of mouse retina tissues, such as an approximately 1.2 times increase in imaging resolution, seamless stitching of single molecules between adjacent optical sections, and a doubling of the sectional interval in volumetric obSTORM imaging due to the extended axial range of usable section thickness. The cubic spline PSF model thus offers a path towards more accurate and faster volumetric obSTORM imaging of biological specimens.