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Authors & Affiliations
Stefanie Kiderlen, Lukas Krainer
Abstract
The experimental requirements for intravital imaging are versatile and varying setups of the microscope are needed. We have developed an integrated imaging platform which gives easy and economically viable access to technically challenging imaging techniques like CARS and SRS, so far typically requiring major system installations and maintenance costs, prohibiting a wider spread to a broader user base. Here, we will demonstrate turn-key portable multimodal multiphoton microscopy, combining 3D label-free upright and inverted imaging that can be integrated into a variety of lab and clinical settings to gain orthogonal informational content, enhance imaging depth, and save time in 3D in vivo and ex vivo samples. The MPX multiphoton microscope allows a large number of different experiments due to its easy-to-use and flexible design providing a large working space underneath the 360-frontend allowing intravital microscopy from any angle and positioning. Multimodal imaging including two-photon (TPEF), higher harmonics (SHG/THG), Coherent anti-Stokes Raman Scattering and Stimulated Raman scattering (CARS/SRS), Fluorescence lifetime (FLIM) and epi-widefield fluorescence gains orthogonal data sets. For deep-tissue imaging in the brain, widefield fluorescence allows functional overview imaging whereas non-linear microscopy enables label-free and deep-tissue intravital imaging in one device! Combined with an ultrafast resonant scanning option even fast dynamic processes like tracking single erythrocytes in brain capillaries can be captured. Future developments will include improvements for even higher imaging resolutions and depth by implementing adaptive optics (AO) techniques and three-photon (3P) microscopy.