Nguyen Trung Duc, Rahmani Amir, Ponjavic Aleks, Millett-Sikking Alfred, Fiolka Reto
Lyda Hill Department of Bioinformatics, UT Southwestern Medical Center, Dallas, TX 75390, USA.
School of Physics and Astronomy, University of Leeds, Leeds LS2 9JT, UK.
Biomed Opt Express. 2025 Feb 26;16(3):1216-1224. doi: 10.1364/BOE.553545. eCollection 2025 Mar 1.
Light-sheet fluorescence microscopy (LSFM) has demonstrated great potential in the life sciences owing to its efficient volumetric imaging capabilities. For long-term imaging, the light-sheet typically needs to be stabilized to the detection focal plane for the best imaging results. Current light-sheet stabilization methods rely on fluorescence emission from the sample, which may interrupt scientific imaging and add to sample photobleaching. Here, we show that for oblique plane microscopes (OPM), a subset of LSFM where a single primary objective is used for illumination and detection, light-sheet stabilization can be achieved without expending sample fluorescence. Our method achieves ∼21 nm axial precision and maintains the light-sheet well within the depth of focus of the detection system for hour-long acquisition runs in a lab environment that would otherwise detune the system. We demonstrate subcellular imaging of the actin skeleton in melanoma cancer cells with a stabilized OPM.
光片荧光显微镜（LSFM）因其高效的体积成像能力在生命科学领域展现出巨大潜力。对于长期成像而言，为获得最佳成像结果，光片通常需要稳定在检测焦平面上。当前的光片稳定方法依赖于样品的荧光发射，这可能会干扰科学成像并加剧样品光漂白。在此，我们表明，对于斜平面显微镜（OPM，LSFM的一个子集，其中单个主物镜用于照明和检测），无需消耗样品荧光即可实现光片稳定。我们的方法实现了约21纳米的轴向精度，并在实验室环境中长达一小时的采集过程中，将光片很好地保持在检测系统的焦深范围内，否则该系统会失调。我们用稳定的OPM展示了黑色素瘤癌细胞中肌动蛋白骨架的亚细胞成像。
