Tissue clearing combined with the use of fluorescent labels has enabled the use of confocal and light sheet microscopy for 3D tissue imaging. However, one of the main issues with many tissue clearing techniques is that the expensive objectives required for deep tissue imaging are not compatible with solvent-based tissue clearing techniques like BABB, 3DISCO, iDISCO, uDISCO and Visikol HISTO which are highly effective. In this publication the researchers show how the use of a corrected multimode fibre (MMF) can achieve deep tissue imaging while preventing damage to the objective. It was shown that the MMF was not constrained by the refractive index of the immersion medium and was able to provide variable working distances. This MMF was used for the fluorescence imaging of beads and stained neuroblastoma cells through optically cleared mouse brain tissue, as well as imaging in an extreme oxidative environment. Additionally, Raman imaging of polystyrene beads was performed in clearing media to demonstrate that this approach may be used for vibrational spectroscopy of optically cleared samples.
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Multimode fibre based imaging for optically cleared samples