Abstract
Motile cilia in the mammalian oviduct play a key role in reproduction, such as transporting fertilized oocytes to the uterus for implantation. Due to their small size (~5–10 μm in length and ~300 nm in diameter), live visualization of cilia and their activity in the lumen of the oviduct through tissue layers represents a major challenge not yet overcome. Here, we report a functional low-coherence optical imaging technique that allows in vivo depth-resolved mapping of the cilia location and cilia beat frequency (CBF) in the intact mouse oviduct with micro-scale spatial resolution. We validate our approach with widely-used microscopic imaging methods, present the first in vivo mapping of the oviduct CBF in its native context, and demonstrate the ability of this approach to differentiate CBF in different locations of the oviduct at different post-conception stages. This technique opens a range of opportunities for live studies in reproductive medicine as well as other areas focused on cilia activity and related ciliopathies.
Highlights
Optical coherence tomography (OCT) is a noninvasive depth-resolved 3D imaging modality that provides micro-scale spatial resolution with the imaging depth of 1–2 mm in highly-scattering tissues[26,27,28]
Our results indicate that described functional OCT (fOCT) approach can be successfully applied to study the dynamic ciliary behavior in mouse oviduct in vivo, opening the door for a variety of live studies of mammalian reproduction and infertility, as well as other research areas involving the analysis of cilia activity
We present and validate an approach for live mapping of cilia and their dynamics in the intact mouse oviduct
Summary
Optical coherence tomography (OCT) is a noninvasive depth-resolved 3D imaging modality that provides micro-scale spatial resolution with the imaging depth of 1–2 mm in highly-scattering tissues[26,27,28]. Focusing on the cilia-driven fluid flow, Jonas, et al and Huang, et al developed OCT-based particle tracking velocimetry to image the quantitative flow dynamics induced by the cilia on Xenopus embryonic model[34,35], which indirectly probes the ciliary function. We combined live mouse manipulation approach with the further development of the OCT-based speckle-variance cilia-detection method[29] to introduce a tomographic imaging technique capable of mapping both the cilia location and the CBF in the intact mouse oviduct in vivo with micro-scale spatial resolution and depth-resolved field of view that covers the whole depth of the oviduct. Our results indicate that described fOCT approach can be successfully applied to study the dynamic ciliary behavior in mouse oviduct in vivo, opening the door for a variety of live studies of mammalian reproduction and infertility, as well as other research areas involving the analysis of cilia activity
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