monitoring-mechanical-evolution-tissue-during-neural-tube-closure

Monitoring the Mechanical Evolution of Tissue During Neural Tube Closure of Chick Embryo

Neural tube closure (NTC) is a critical process during embryonic development. Failure in this process can lead to neural tube defects, causing congenital ...

Wayne State University

To understand the dynamics of tissue stiffness during neural tube formation and closure in a murine model, we have developed a multimodal, coaligned imaging system combining optical coherence tomography (OCT) and Brillouin microscopy. Brillouin microscopy can map the longitudinal modulus of tissue but cannot provide structural images. Thus, it is limited for imaging dynamic processes such as neural tube formation and closure. To overcome this limitation, we have combined Brillouin microscopy and OCT in one coaligned instrument. OCT provided depth-resolved structural imaging with a micrometer-scale spatial resolution to guide stiffness mapping by Brillouin modality. 2D structural and Brillouin frequency shift maps were acquired of mouse embryos at gestational day (GD) 8.5, 9.5, and 10.5 with the multimodal system. The results demonstrate the capability of the system to obtain structural and stiffness information simultaneously.

Multimodal imaging system combining optical coherence tomography and Brillouin microscopy for neural tube imaging.

Neural tube closure (NTC) is a complex process of embryonic development involving molecular, cellular, and biomechanical mechanisms. While the genetic factors and biochemical signaling have been extensively investigated, the role of tissue biomechanics remains mostly unexplored due to the lack of tools. Here, we developed an optical modality that can conduct time-lapse mechanical imaging of neural plate tissue as the embryo is experiencing neurulation. This technique is based on the combination of a confocal Brillouin microscope and a modified ex ovo culturing of chick embryo with an on-stage incubator. With this technique, for the first time, we captured the mechanical evolution of the neural plate tissue with live embryos. Specifically, we observed the continuous increase in tissue modulus of the neural plate during NTC for ex ovo cultured embryos, which is consistent with the data of in ovo culture as well as previous studies. Beyond that, we found that the increase in tissue modulus was highly correlated with the tissue thickening and bending. We foresee this non-contact and label-free technique opening new opportunities to understand the biomechanical mechanisms in embryonic development.

Time-lapse mechanical imaging of neural tube closure in live embryo using Brillouin microscopy.

Brillouin microscopy is a technique for mechanical characterization of biological material without contact at high three-dimensional resolution. Here, we introduce dual line-scanning Brillouin microscopy (dLSBM), which improves acquisition speed and reduces irradiation dose by more than one order of magnitude with selective illumination and single-shot analysis of hundreds of points along the incident beam axis. Using tumor spheroids, we demonstrate the ability to capture the sample response to rapid mechanical perturbations as well as the spatially resolved evolution of the mechanical properties in growing spheroids.

Rapid biomechanical imaging at low irradiation level via dual line-scanning Brillouin microscopy.

Multimodal optical imaging techniques are useful for various applications, including imaging biological samples for providing comprehensive material properties. In this work, we developed a new modality that can measure a set of mechanical, optical, and acoustical properties of a sample at microscopic resolution, which is based on the integration of Brillouin (Br) and photoacoustic (PA) microscopy. The proposed multimodal imaging technique not only can acquire co-registered Br and PA signals but also allows us to utilize the sound speed measured by PA to quantify the sampleâ€™s refractive index, which is a fundamental property of the material and cannot be measured by either technique individually. We demonstrated the colocalization of Br and time-resolved PA signals in a synthetic phantom made of kerosene and CuSO  aqueous solution. In addition, we measured the refractive index of saline solutions and validated the result against published data with a relative error of 0.3 %. This multimodal Br-PA modality could open a new way for characterizing biological samples in physiological and pathological conditions.

A versatile multimodal optical modality based on Brillouin light scattering and photoacoustic effect.

Publisher Correction: Rapid biomechanical imaging at low irradiation level via dual line-scanning Brillouin microscopy.

To elucidate the physiological, cellular, and molecular mechanisms responsible for initiating and sustaining ocular neuropathic pain, we created a blue-light-exposure model in C57BL/6 mice. Mice were exposed to 12 h of blue or white light followed by 12 h of darkness. Before blue light exposure, baseline tear secretion, stability, and ocular hyperalgesia were assessed by measuring hyper- or hypo-osmotic solution-induced eye wiping, wind-induced eye closing, and cold-induced eye blinking. At 1 day post-blue light exposure, alterations in hypotonic/hypertonic-induced eye-wiping, and tear film abnormalities were observed. Eye-wiping behaviors were abolished by topical anesthesia. The cold-stimulated eye-blinking and wind-stimulated eye-closing behaviors began after day 3 and their frequency further increased after day 9. Blue-light exposure reduced the density of nerve endings, and increased their tortuosity, the number of beadlike structures, and the branching of stromal nerve fibers, as assessed by whole-mount confocal microscopy. Blue-light exposure also increased TRPV1, but not TRPV4 staining intensity of corneal-projecting neurons in the trigeminal ganglia, as detected by Fluorogold retrograde labeling and immunohistochemistry. TRPV1 and substance P expression was increased, whereas CGRP expression deceased at the mRNA level in isolated corneal projecting neurons. Hence, our blue-light exposure B6 mouse model for assessing tearing and ocular hyperalgesia is useful for studying ocular pain and its underlying mechanisms. Blue-light-induced alterations in tearing and ocular hyperalgesia may be related to the elevated expression of TRPV1, SP, and/or the suppressed expression of CGRP at the ocular surface.

Ocular nociception and neuropathic pain initiated by blue light stress in C57BL/6J mice.

Brillouin microscopy based on spontaneous Brillouin scattering has emerged as a unique elastography technique because of its merit of non-contact, label-free, and high-resolution mechanical imaging of biological cell and tissue. Recently, several new optical modalities based on stimulated Brillouin scattering have been developed for biomechanical research. As the scattering efficiency of the stimulated process is much higher than its counterpart in the spontaneous process, stimulated Brillouin-based methods have the potential to significantly improve the speed and spectral resolution of existing Brillouin microscopy. Here, we review the ongoing technological advancements of three methods, including continuous wave stimulated Brillouin microscopy, impulsive stimulated Brillouin microscopy, and laser-induced picosecond ultrasonics. We describe the physical principle, the representative instrumentation, and biological application of each method. We further discuss the current limitations as well as the challenges for translating these methods into a visible biomedical instrument for biophysics and mechanobiology.

Non-contact and label-free biomechanical imaging: Stimulated Brillouin microscopy and beyond.

Epithelial-mesenchymal transition (EMT) is a biological process where epithelial cells lose their adhesive properties and gain invasive, metastatic, and mesenchymal properties. Maintaining the balance between the epithelial and mesenchymal stage is essential for tissue homeostasis. Many of the genes promoting mesenchymal transformation have been identified; however, our understanding of the genes responsible for maintaining the epithelial phenotype is limited. Our objective was to identify the genes responsible for maintaining the epithelial phenotype and inhibiting EMT.

Article	Year
Multimodal imaging system combining optical coherence tomography and Brillouin microscopy for neural tube imaging. Optics letters\| PubMed ID: 35290310	2022
Time-lapse mechanical imaging of neural tube closure in live embryo using Brillouin microscopy. Scientific reports\| PubMed ID: 36609620	2023
Rapid biomechanical imaging at low irradiation level via dual line-scanning Brillouin microscopy. Nature methods\| PubMed ID: 36894684	2023
A versatile multimodal optical modality based on Brillouin light scattering and photoacoustic effect. bioRxiv : the preprint server for biology\| PubMed ID: 36945550	2023
Publisher Correction: Rapid biomechanical imaging at low irradiation level via dual line-scanning Brillouin microscopy. Nature methods\| PubMed ID: 37069272	2023
Ocular nociception and neuropathic pain initiated by blue light stress in C57BL/6J mice. Pain\| PubMed ID: 37093736	2023
Non-contact and label-free biomechanical imaging: Stimulated Brillouin microscopy and beyond. Frontiers in physics\| PubMed ID: 37377499	2023
A Novel Role of Connective Tissue Growth Factor in the Regulation of the Epithelial Phenotype. Cancers\| PubMed ID: 37835529	2023

Jitao Zhang

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