On-Axis 2-Photon Light-Sheet Generation for a 3-D in-vivo Imaging (Microscope) System (OASIS)

Eurostars Project E!10266


Subcellular dynamics in intact tissue is usually being studied using 2-photon-pointscanning or light-sheet microscopy. The former excels in penetration depth, but lacks speed and doesn’t preserve cell viability as well as the latter, which suffers from a limited z-resolution and requires tedious tissue mounting procedures. With OASIS we will retain the inline geometry of the classical microscope, boost its speed beyond that of a light-sheet microscope and provide a significantly better z-resolution.

Working Plan

We will build and characterize a novel, patent-pending 2-photon (2P) light-sheet microscope for minimally invasive sub-cellular imaging of live tissue. Its optical sectioning capabilities, large field-of-view, high speed and low photodamage are achieved by using spatially and temporally changing 2P excitation to create a light sheet. Fluorescence is confocally imaged through the same objective onto a fast camera. The concept will be validated by imaging brain astroglial Ca2+ signals.

Goal and Perspectives

The project goal is to establish a new technology, which has the potential of opening a whole new market segment for in vivo live-cell imaging. We will develop a prototype that will serve to validate the concept, compare it to existing technologies and prove its virtues with respect to cell viability. The new technology will be integrated in a stand-alone upright microscope, driven by proprietary control electronics and software. The design will be modular, so that it will also be possible to market an attachment, based on the same technology, to existing microscopes.