Tetrahedral serial multiview microscopy and image fusion for improved resolution and extent in stained zebrafish embryos.
Tetrahedral serial multiview microscopy and image fusion for improved resolution and extent in stained zebrafish embryos.
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四面体串行多视图显微镜和图像融合可提高染色斑马鱼胚胎的分辨率和范围。
DOI：
10.1002/dvdy.683
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Megason,SeanG
中科院分区：
文献类型：
--
作者：
Kroll,JohannaB;Cha,Anna;Oyler-Yaniv,Alon;Lambert,Talley;Swinburne,IanA;Murphy,Andrew;Megason,SeanG
背景在整个生物体的单细胞水平上的空间映射可以揭示参与脊椎动物发育的分子角色。已经开发出使用多个物镜同时从多个视图观察样品的定制显微镜。这种多视图成像方法可以提高分辨率和图像质量的均匀性，并且允许对整个胚胎进行成像（Swoger等人，Opt Express，2007;15（13）：8029）。然而，多视图成像是高度受限的专业设备，需要多个物镜或样品旋转与自动化hardware.ResultsOur方法使用一个标准的单物镜共聚焦显微镜进行串行多视图成像。通过将固定的样品安装在琼脂糖四面体中顺序地对多个视图进行成像，所述琼脂糖四面体在成像每个面之间手动旋转。计算图像融合允许从不同角度采集的多个平铺Z堆栈创建联合3D图像。融合后的图像具有更高的分辨率和成像范围。ConclusionWith this technique，multiview imaging can be performed on a variety of common single‐objective microscopes to allow for whole‐embryo，high-resolution imaging.
BackgroundSpatial mapping on the single‐cell level over the whole organism can uncover roles of molecular players involved in vertebrate development. Custom microscopes have been developed that use multiple objectives to view a sample from multiple views at the same time. Such multiview imaging approaches can improve resolution and uniformity of image quality as well as allow whole embryos to be imaged (Swoger et al.,Opt Express, 2007;15(13):8029). However, multiview imaging is highly restricted to specialized equipment requiring multiple objectives or sample rotation with automated hardware.ResultsOur approach uses a standard single‐objective confocal microscope to perform serial multiview imaging. Multiple views are imaged sequentially by mounting the fixed sample in an agarose tetrahedron that is manually rotated in between imaging each face. Computational image fusion allows for a joint 3D image to be created from multiple tiled Z‐stacks acquired from different angles. The resulting fused image has improved resolution and imaging extent.ConclusionWith this technique, multiview imaging can be performed on a variety of common single‐objective microscopes to allow for whole‐embryo, high‐resolution imaging.
DOI：
10.3791/53966
发表时间：
2016-04-10
期刊：
Journal of visualized experiments : JoVE
影响因子：
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作者：
Icha J;Schmied C;Sidhaye J;Tomancak P;Preibisch S;Norden C
通讯作者：
Norden C
影响因子：
4.6
作者：
Shkumatava, A;Fischer, S;Neumann, CJ
通讯作者：
Neumann, CJ
影响因子：
64.5
作者：
Munjal A;Hannezo E;Tsai TY;Mitchison TJ;Megason SG
通讯作者：
Megason SG