Optical sectioning deep inside live embryos by selective plane illumination microscopy
Optical sectioning deep inside live embryos by selective plane illumination microscopy
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DOI：
10.1126/science.1100035
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发表时间：
2004-08-13
期刊：
影响因子：
56.9
通讯作者：
Stelzer, EHK
中科院分区：
文献类型：
--
作者：
Huisken, J;Swoger, J;Stelzer, EHK
大型活体生物样本因其吸收和散射特性而对现有光学成像技术提出了挑战。我们开发了选择性平面照明显微镜 (SPIM)，可以生成尺寸达几毫米的样品的多维图像。该系统将二维照明与基于正交相机的检测相结合，以实现整个样品的高分辨率光学切片成像，光损伤最小，并且能够以能够捕获瞬态生物现象的速度。我们使用 SPIM 对转基因青鳉品系 Arnie 体内的所有肌肉进行可视化，该品系在肌肉组织中表达绿色荧光蛋白。我们还证明 SPIM 可用于可视化相对不透明的果蝇体内的胚胎发生。
Large, living biological specimens present challenges to existing optical imaging techniques because of their absorptive and scattering properties. We developed selective plane illumination microscopy (SPIM) to generate multidimensional images of samples up to a few millimeters in size. The system combines two-dimensional illumination with orthogonal camera-based detection to achieve high-resolution, optically sectioned imaging throughout the sample, with minimal photodamage and at speeds capable of capturing transient biological phenomena. We used SPIM to visualize all muscles in vivo in the transgenic Medaka line Arnie, which expresses green fluorescent protein in muscle tissue. We also demonstrate that SPIM can be applied to visualize the embryogenesis of the relatively opaque Drosophila melanogaster in vivo.