The TriScan: fast and sensitive 3D confocal fluorescence imaging using a simple optical design

Abstract

AbstractWe present the TriScan, a compact and inexpensive fluorescence microscope that combines the speed of widefield microscopy with the 3D-sectioning capabilities of confocal microscopy. The optical layout is based on an add-on module that combines line-scan confocal imaging with a sensitive camera detector, realized using a simple optical layout that permits the use of arbitrarily fast scanning mirrors. The resulting design is theoretically capable of full field-of-view acquisition rates in the kilohertz regime combined with a diffraction-limited resolution and single-molecule sensitivity. Overall, the TriScan microscope provides the ease-of-use and speed of widefield imaging combined with the optical sectioning of one-photon confocal imaging, in a simple and inexpensive design suitable for a broad variety of settings ranging from research to diagnostic applications and screening.This bioRxiv manuscript describes an ongoing research project and associated preliminary data acquired using an early prototype of the instrument. We welcome and appreciate your enquiries, suggestions, and feedback. Updated versions of this manuscript will be deposited as the project progresses.The author list reflects the core team and points of contact working on this project, but does not reflect all of the contributions made to this research thus far. We are particularly grateful to Damla Temel (formerly KU Leuven) for assistance in the construction of the initial prototype, Lydia Danglot (IPNP Paris) and Hugo Vankelecom (KU Leuven) for providing samples, and Marcel Leutenegger (Max Planck Institute for Biophysical Research) for initial discussions regarding the practical implementation. We also thank the Research Foundation-Flanders (FWO) and the European Research Council for financial support.