Multiplexed PSF engineering for 3D multicolor particle tracking

TL;DR

This paper presents a multiplexed PSF engineering method enabling efficient 3D multicolor particle tracking across large fields of view, overcoming previous trade-offs in optical imaging.

Contribution

It introduces a novel multiplexed PSF engineering technique that allows simultaneous 3D multicolor tracking with high photon efficiency and large field of view.

Findings

Successfully tracked five emitter types in vitro

Achieved co-localization of DNA loci in live yeast cells

Demonstrated large field of view and spectral specificity

Abstract

Three-dimensional spatiotemporal tracking of microscopic particles in multiple colors is a challenging optical imaging task. Existing approaches require a trade-off between photon-efficiency, field of view, mechanical complexity, spectral specificity and speed. Here, we introduce multiplexed point-spread function engineering that achieves photon efficient, 3D, multicolor particle tracking over a large field of view. This is accomplished by first chromatically splitting the emission path of a microscope to different channels, engineering the point-spread function of each, and then recombining them onto the same region of the camera. We demonstrate our technique for simultaneously tracking five types of emitters in-vitro, as well as co-localization of DNA loci in live yeast cells.