Gusachenko Ivan, Nylk Jonathan, Tello Javier A, Dholakia Kishan
SUPA, School of Physics and Astronomy, University of St. Andrews, Fife, KY16 9SS, UK.
School of Medicine, University of St. Andrews, Fife, KY16 9TF, UK.
Biomed Opt Express. 2017 Oct 23;8(11):5179-5190. doi: 10.1364/BOE.8.005179. eCollection 2017 Nov 1.
Optical clearing is emerging as a popular approach particularly for studies in neuroscience. However the use of corrosive clearing solutions typically requires sophisticated objectives or extreme care with optical components chosen for single- or multi-photon imaging. In contrast to the use of complex, custom-made microscope objectives, we show that the use of a corrected multimode fibre (MMF) offers a route that is resistant to corrosion, can be used in clearing media, is not constrained by the refractive index of the immersion medium and offers flexible working distances. Using a corrected MMF, we demonstrate fluorescence imaging of beads and stained neuroblastoma cells through optically cleared mouse brain tissue, as well as imaging in an extreme oxidative environment to show the versatility of our approach. Additionally, we perform Raman imaging of polystyrene beads in clearing media to demonstrate that this approach may be used for vibrational spectroscopy of optically cleared samples.
光学透明化正成为一种流行的方法,特别是在神经科学研究中。然而,使用腐蚀性透明化溶液通常需要精密的物镜,或者在为单光子或多光子成像选择光学组件时要格外小心。与使用复杂的定制显微镜物镜不同,我们表明使用校正多模光纤(MMF)提供了一条抗腐蚀的途径,可用于透明化介质中,不受浸没介质折射率的限制,并提供灵活的工作距离。使用校正的MMF,我们展示了通过光学透明化的小鼠脑组织对珠子和染色的神经母细胞瘤细胞进行荧光成像,以及在极端氧化环境中成像,以展示我们方法的多功能性。此外,我们对透明化介质中的聚苯乙烯珠子进行拉曼成像,以证明这种方法可用于光学透明化样品的振动光谱分析。