University of Gothenburg, Biomedical Photonics Group, Department of Chemistry and Molecular Biology,, Sweden.
University of Gothenburg, Department of Chemistry and Molecular Biology, Faculty of Science, Gothenb, Sweden.
J Biomed Opt. 2020 May;25(7):1-11. doi: 10.1117/1.JBO.25.7.071205.
Research in tissue engineering and in vitro organ formation has recently intensified. To assess tissue morphology, the method of choice today is restricted primarily to histology. Thus novel tools are required to enable noninvasive, and preferably label-free, three-dimensional imaging that is more compatible with futuristic organ-on-a-chip models.
We investigate the potential for using multiphoton microscopy (MPM) as a label-free in vitro approach to monitor calcium-induced epidermal differentiation.
In vitro epidermis was cultured at the air-liquid interface in varying calcium concentrations. Morphology and tissue architecture were investigated using MPM based on visualizing cellular autofluorescence.
Distinct morphologies corresponding to epidermal differentiation were observed. In addition, Ca2 + -induced effects could be distinguished based on the architectural differences in stratification in the tissue cultures.
Our study shows that MPM based on cellular autofluorescence enables visualization of Ca2 + -induced differentiation in epidermal skin models in vitro. The technique has potential to be further adapted as a noninvasive, label-free, and real-time tool to monitor tissue regeneration and organ formation in vitro.
组织工程和体外器官形成的研究最近得到了加强。为了评估组织形态，目前首选的方法主要局限于组织学。因此，需要新的工具来实现非侵入性的、最好是无标记的三维成像，这更符合未来的器官芯片模型。
我们研究了多光子显微镜（MPM）作为一种无标记的体外方法，用于监测钙诱导的表皮分化的潜力。
在不同钙离子浓度下，将体外表皮在气液界面上培养。使用基于细胞自发荧光可视化的 MPM 研究形态和组织结构。
观察到与表皮分化相对应的不同形态。此外，还可以根据组织培养中分层的结构差异，区分 Ca2+诱导的效应。
我们的研究表明，基于细胞自发荧光的 MPM 能够可视化体外表皮皮肤模型中 Ca2+诱导的分化。该技术有可能进一步被改编为一种非侵入性、无标记和实时工具，以监测体外组织再生和器官形成。
