Localization precision in chromatic multifocal imaging

Abstract

Multifocal microscopy affords fast acquisition of microscopic 3D images. This is made possible using a multifocal grating optic, however this induces chromatic dispersion effects into the point spread function impacting image quality and single-molecule localization precision. To minimize this effect, researchers use narrow-band emission filters. However, the choice of optimal emission filter bandwidth in such systems is, thus far, unclear. This work presents a theoretical framework to investigate how the localization precision of a point emitter is affected by the emission filter bandwidth. We calculate the Cram\'er-Rao lower bound for the 3D position of a single emitter imaged using a chromatic multifocal microscope. Results show that the localization precision improves with broader emission filter bandwidth due to increased photon throughput, despite a larger chromatic dispersion. This study provides a framework for optimally designing chromatic multifocal optics and serves as a theoretical foundation for interpretting results.