Despite their tremendous contribution to biomedical research and diagnosis, conventional spatial sampling techniques such as wide-field, point scanning or selective plane illumination microscopy face inherent limiting trade-offs between spatial resolution, field-of-view, phototoxicity and recording speed. Several of these trade-offs are the result of spatial sampling with diffracting beams. Here, we introduce a new strategy for fluorescence imaging, SWIF, which instead encodes the axial profile of a sample in the Fourier domain. We demonstrate how this can be achieved with propagation-invariant illumination patterns that extend over several millimeters and robustly propagate through layers of varying refractive index. This enabled us to image a lateral field-of-view of 0.8 mm x 1.5 mm with an axial resolution of 2.4 µm – greatly exceeding the lateral field-of-view of conventional illumination techniques (~100 µm) at comparable resolution. Thus, SWIF allowed us to surpass the limitations of diffracting illumination beams and untangle lateral field-of-view from resolution.
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机译:尽管它们对生物医学研究和诊断做出了巨大贡献,但传统的空间采样技术(例如宽视场,点扫描或选择性平面照明显微镜)在空间分辨率,视野,光毒性和记录速度之间面临着固有的局限性折衷。这些折衷中的几个是使用衍射光束进行空间采样的结果。在这里,我们介绍了一种新的荧光成像策略,即SWIF,它可以在傅立叶域中编码样本的轴向轮廓。我们演示了如何使用传播不变的照明图案来实现这一目标,该照明图案延伸超过几毫米,并牢固地传播通过折射率不同的层。这使我们能够以2.4 µm的轴向分辨率对0.8 mm x 1.5 mm的横向视场成像–在相当的分辨率下大大超过了传统照明技术(〜100 µm)的横向视场。因此,SWIF允许我们克服衍射光束的限制,并从分辨率上解开横向视场。
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