In vivo micro-scale tomography of ciliary behavior in the mammalian oviduct

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.

Dynamic imaging and quantitative analysis of cranial neural tube closure in the mouse embryo using optical coherence tomography

“…Details of the OCT system were described in our previous work [56]. Briefly, the system employs a near-infrared laser with ~808 nm central wavelength and ~110 nm bandwidth.…”

Ex vivo visualization of human ciliated epithelium and quantitative analysis of induced flow dynamics by using optical coherence tomography

“…However, it was possible to utilize the temporal frequency analysis to spectrally filter the signal and reject the noises from the mucus. As we presented in Figure , the ciliary activities induced dominant frequencies in the temporal frequency domain, and we could use both amplitude and frequency of the dominant frequencies to differentiate the cilia against the mucus . It is worth mentioning that the mucus located in the proximity of the ciliated epithelium possessed a similar dominant frequency as the underlying cilia.…”

“…Previously, this parameter was mainly manually measured even with the aid of OCT . Recently, temporal frequency/Fourier analysis was adopted by different groups on different subjects, including mice oviduct and swine trachea . In this manuscript, we presented a similar demonstration on ex vivo human trachea.…”

Metabolite availability as a window to view the early embryo microenvironment in vivo

“…More studies should seek to eavesdrop on the communication between the embryo and maternal epithelium, which should enable us to define new metrics for improving the implantation rates and pregnant outcome following in vitro embryo culture. The progress made in live‐cell imaging and digital videomicroscopy should also allow us to study physiological and pathological events in the oviduct under near‐in vivo conditions, further revealing what happens in the female tract (Kolle, ; Wang, Burton, Behringer, & Larina, ). Such approaches will allow us to create systems wherein the preimplantation embryo can “return” to the in vivo environment (Menezo et al ().…”

“…Could such physiologic variation explain why human embryos cultured in human tubal fluid in vitro show lower implantation rate and/or live birth rate than those cultured in a synthetic G5 medium (Kleijkers et al, 2015(Kleijkers et al, , 2016Mantikou et al, 2016)? further revealing what happens in the female tract (Kolle, 2012;Wang, Burton, Behringer, & Larina, 2015). Such approaches will allow us to create systems wherein the preimplantation embryo can "return" to the in vivo environment (Menezo et al (2015).…”