Increasing the acquisition speed in oblique plane microscopy via aliasing.

Abstract

Oblique plane microscopy (OPM), a variant of light-sheet fluorescence microscopy (LSFM), enables rapid volumetric imaging without mechanically scanning the sample or an objective. In an OPM, the sample space is mapped to a distortion-free image space via remote focusing, and the oblique light-sheet plane is mapped onto a camera via a tilted tertiary imaging system. As a result, the 3D point-spread function and optical transfer function (OTF) are tilted to the optical axis of the tertiary imaging system. To satisfy Nyquist sampling, small scanning steps are required to encompass the tilted 3D OTF, slowing down acquisition and increasing sample exposure. Here, we show that a judicious amount of under-sampling can lead to a form of aliasing in OPM that can be recovered without a loss of spatial resolution while minimizing artifacts. The resulting speed gains depend on the optical parameters of the system and reach 2-4-fold in our demonstrations. We leverage this method for rapid subcellular 3D imaging of mitochondria and the endoplasmic reticulum.