Microscopy seeks to simultaneously maximize optical resolution, contrast, speed, volume size, and probe tolerability, which is possible by combining different complementary imaging techniques with their specific strengths. Here, we show how to combine three physical concepts to increase resolution and contrast in light-sheet microscopy by making the effective light-sheet thinner through phase shaping, fluorophores-switching, and dynamic blocking of fluorescence. This shape-switch-block principle is realized by illumination with two holographically shaped, sectioned Bessel beams. Second, by switching off fluorophores in the proximity of the excitation center using continuous-wave stimulated emission depletion (STED). And third, by blocking fluorescence outside the switching region by confocal line detection. Thereby, we reduce the light-sheet thickness by 35%, achieving an isotropic resolution with beads in a 300 × 70 × 50 µm³ volume. Without STED, we obtain 0.37 µm resolution in cell clusters at improved sectioning and penetration depth. The shape-switch-block concept promises high potential, also for other microscopy techniques.

显微镜试图同时使光学分辨率，对比度，速度，体积大小和探针耐受性最大化，这可以通过将不同的互补成像技术及其特定强度相结合来实现。在这里，我们展示了如何通过相整形，荧光团转换和动态荧光阻断使有效的光片变薄，从而结合三种物理概念来提高光片显微镜的分辨率和对比度。这种形状切换块原理是通过用两个全息形状的分段贝塞尔光束进行照明来实现的。第二，通过使用连续波激发的发射损耗（STED）关闭激发中心附近的荧光团。第三，通过共焦线检测阻断开关区域外的荧光。从而，我们将光片的厚度减少了35％，以300×70×50 µm³体积的珠子实现了各向同性分辨率。如果不使用STED，我们可以在改进的切片和穿透深度下获得0.37 µm的细胞簇分辨率。形状切换块概念也有望在其他显微镜技术中发挥出巨大的潜力。