Cell Biology and Biophysics Unit, European Molecular Biology Laboratory, Heidelberg, Germany.
Shanghai Institute of Optics and Fine Mechanics, Chinese Academy of Sciences, Shanghai, China.
Nat Methods. 2023 Dec;20(12):1971-1979. doi: 10.1038/s41592-023-02054-z. Epub 2023 Oct 26.
Brillouin microscopy is an emerging optical elastography technique capable of assessing mechanical properties of biological samples in a three-dimensional, all-optical and noncontact fashion. The typically weak Brillouin scattering signal can be substantially enhanced via a stimulated Brillouin scattering (SBS) process; however, current implementations require high pump powers, which prohibit applications to photosensitive or live imaging of biological samples. Here we present a pulsed SBS scheme that takes advantage of the nonlinearity of the pump-probe interaction. In particular, we show that the required pump laser power can be decreased ~20-fold without affecting the signal levels or spectral precision. We demonstrate the low phototoxicity and high specificity of our pulsed SBS approach by imaging, with subcellular detail, sensitive single cells, zebrafish larvae, mouse embryos and adult Caenorhabditis elegans. Furthermore, our method permits observing the mechanics of organoids and C. elegans embryos over time, opening up further possibilities for the field of mechanobiology.
布里渊显微镜是一种新兴的光学弹性成像技术,能够以三维、全光学和非接触的方式评估生物样本的力学性能。典型的弱布里渊散射信号可以通过受激布里渊散射(SBS)过程得到显著增强;然而,当前的实现方法需要高泵浦功率,这限制了其在生物样本的光敏感或活细胞成像中的应用。在这里,我们提出了一种利用泵浦-探测相互作用的非线性的脉冲 SBS 方案。具体来说,我们表明,在不影响信号水平或光谱精度的情况下,可以将所需的泵浦激光功率降低约 20 倍。我们通过对敏感的单细胞、斑马鱼幼虫、小鼠胚胎和成年秀丽隐杆线虫进行亚细胞细节成像,展示了我们的脉冲 SBS 方法的低光毒性和高特异性。此外,我们的方法允许观察类器官和秀丽隐杆线虫胚胎的力学特性随时间的变化,为机械生物学领域开辟了更多的可能性。