RT Journal Article
SR Electronic
T1 Multi-photon attenuation-compensated light-sheet fluorescence microscopy
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2019.12.12.873877
DO 10.1101/2019.12.12.873877
A1 Madhu Veettikazhy
A1 Jonathan Nylk
A1 Federico Gasparoli
A1 Adrià Escobet-Montalbán
A1 Anders Kragh Hansen
A1 Dominik Marti
A1 Peter Eskil Andersen
A1 Kishan Dholakia
YR 2019
UL http://biorxiv.org/content/early/2019/12/12/2019.12.12.873877.abstract
AB Attenuation of optical fields owing to scattering and absorption limits the penetration depth for imaging. Whilst aberration correction may be used, this is difficult to implement over a large field-of-view in heterogeneous tissue. Attenuation-compensation allows tailoring of the maximum lobe of a propagation-invariant light field and promises an increase in depth penetration for imaging. Here we show this promising approach may be implemented in multi-photon (two-photon) light-sheet fluorescence microscopy and, furthermore, be achieved in a facile manner utilizing a graded neutral density filter, circumventing the need for complex beam shaping apparatus. A “gold standard” system utilizing a spatial light modulator for beam shaping is used to benchmark our implementation. The approach will open up enhanced depth penetration in light-sheet imaging to a wide range of end users.