Beyond comparison: Brillouin microscopy and AFM-based indentation reveal divergent insights into the mechanical profile of the murine retina

IF 4.6 Q1 OPTICS Journal of Physics-Photonics Pub Date : 2024-07-01 DOI:10.1088/2515-7647/ad5ae3
Marcus Gutmann, Jana Bachir Salvador, Paul Müller, Kyoohyun Kim, Martin Schicht, Serhii Aif, Friedrich Paulsen, Lorenz Meinel, Jochen Guck and Stephanie Möllmert
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Abstract

Mechanical tissue properties increasingly serve as pivotal phenotypic characteristics that are subject to change during development or pathological progression. The quantification of such material properties often relies on physical contact between a load-applying probe and an exposed sample surface. For most tissues, meeting these requirements entails an invasive preparation, which poses the risk of yielding mechanical properties that do not portray the physiological state of a tissue within a functioning organism. Brillouin microscopy has emerged as a non-invasive, optical technique that enables the assessment of mechanical cell and tissue properties with high spatio-temporal resolution. In optically transparent specimens, it does not require animal sacrifice, tissue dissection or sectioning. However, the extent to which results obtained from Brillouin microscopy allow to infer conclusions about potential results obtained with a contact-based technique, and vice versa, is unclear. Sources for discrepancies include the varying characteristic temporal and spatial scales, the directionality of measurement, environmental factors, and mechanical moduli probed. In this work, we addressed those aspects by quantifying the mechanical properties of acutely dissected murine retinae using Brillouin microscopy and atomic force microscopy (AFM)-based indentation measurements. Our results show a distinct mechanical profile of the retinal layers with respect to the Brillouin frequency shift, the Brillouin linewidth and the apparent Young’s modulus. Contrary to previous reports, our findings do not support a simple correlative relationship between Brillouin frequency shift and apparent Young’s modulus. Additionally, the divergent sensitivities of Brillouin microscopy and AFM-indentation measurements to structural features, as visualized by transmission electron microscopy, to cross-linking or changes post mortem underscore the dangers of assuming interchangeability between the two methods. In conclusion, our study advocates for viewing Brillouin microscopy and AFM-based indentation measurements as complementary tools, discouraging direct comparisons a priori and suggesting their combined use for a more comprehensive understanding of tissue mechanical properties.
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超越比较:布里渊显微镜和基于原子力显微镜的压痕法揭示了小鼠视网膜机械特征的不同见解
组织的机械特性越来越成为关键的表型特征,在发育或病理发展过程中会发生变化。此类材料特性的量化通常依赖于加载探针与暴露样本表面之间的物理接触。对于大多数组织来说,要满足这些要求就必须进行侵入性制备,这就有可能导致所获得的机械特性无法反映正常机体内组织的生理状态。布里渊显微镜是一种非侵入式光学技术,能以高时空分辨率评估细胞和组织的机械特性。在光学透明的标本中,布里渊显微镜无需牺牲动物、解剖组织或切片。然而,从布里渊显微镜获得的结果能在多大程度上推断出接触式技术可能获得的结果,反之亦然,这一点尚不清楚。造成差异的原因包括不同的特征时空尺度、测量的方向性、环境因素和探测的机械模量。在这项工作中,我们利用布里渊显微镜和基于原子力显微镜(AFM)的压痕测量方法量化了急性剥离的鼠视网膜的机械特性,从而解决了这些方面的问题。我们的结果表明,视网膜层在布里渊频移、布里渊线宽和表观杨氏模量方面具有独特的力学特征。与之前的报告相反,我们的研究结果并不支持布里渊频移与表观杨氏模量之间的简单相关关系。此外,布里渊显微镜和原子力显微镜-压痕测量法对透射电子显微镜观察到的结构特征、交联或死后变化的敏感性不同,这突出表明假设这两种方法可以互换的危险性。总之,我们的研究主张将布里渊显微镜和原子力显微镜压痕测量视为互补工具,不鼓励先验地进行直接比较,而是建议将两者结合使用,以便更全面地了解组织的机械特性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
CiteScore
10.70
自引率
0.00%
发文量
27
审稿时长
12 weeks
期刊最新文献
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