Buchanan, JoAnn (Stanford) (specimen prep), Genoud, Christel (Gatan) (imaging) (2010) CIL:6229, Danio rerio. CIL. Dataset

This study on the zebrafish optic tectum is a series of four images with magnifications ranging from the SEM block face overview to of the zebrafish head to high magnification (80,000X) showing optic tectum synapse.
The first image is a bloc face SEM image at 200x of the epon embedded zebrafish head. This image, the second image in the series, is another block face SEM low magnification image of the tectum. The third image is a 60,000X TEM image of a OTO fixation and copper lead en bloc staining of zebrafish optic tectum synapse. The fourth image is a 80,000X TEM image of a zebrafish optic tectum synapse.

This low magnification image larval zebrafish image of the optic tectum was collected on a Gatan 3View® serial block face scanning electron microscope (SBFSEM). The 3View® is attached to a FEI Quanta 600 FEG SEM. Block face imaging is a way to look at epon embedded specimens using backscattered electrons in the SEM. A diamond knife in the vacuum chamber of the SEM cuts hundreds of ultrathin serial sections, thus gathering a perfected registered stack of images.

Reference for SBFSEM: Denk, W. and H. Horstmann. Serial block face scanning electron microscopy to reconstruct three- dimensional tissue nanostructure. PloS Biology vol 2, 11; e329 doi:10.1371/journal.pbio.0020329

Microwave processing, osmium tetroxoxide/osmid procedure for enhancing membranes, and copper lead en bloc staining.
Procedure for specimen preparation available at http://www.stanford.edu/~redhair/JoAnn_Buchanan.

本项关于斑马鱼视觉顶盖的研究包含四幅图像，其放大倍数从扫描电子显微镜（SEM）的块状面概览至斑马鱼头部的高倍放大（80,000倍），展示了视觉顶盖的突触。首幅图像是200倍放大倍数的块状面SEM图像，展示了环氧树脂嵌入的斑马鱼头部。此图像为系列中的第二幅图像，是另一幅低倍放大的块状面SEM图像，展示了顶盖。第三幅图像为60,000倍透射电子显微镜（TEM）图像，展示了斑马鱼视觉顶盖突触的OTO固定和铜铅块状染色。第四幅图像为80,000倍TEM图像，展示了斑马鱼视觉顶盖突触。此低倍放大图像的斑马鱼幼虫视觉顶盖图像是在Gatan 3View®连续块状面扫描电子显微镜（SBFSEM）上收集的。3View®连接到FEI Quanta 600 FEG SEM。块状面成像是一种利用SEM中背散射电子观察环氧树脂嵌入标本的方法。SEM真空室中的钻石刀切割数百个超薄连续切片，从而收集到一个完美注册的图像堆栈。关于SBFSEM的参考文献：Denk, W. 和 H. Horstmann. 序列块状面扫描电子显微镜用于重建三维组织纳米结构。PloS Biology vol 2, 11; e329 doi:10.1371/journal.pbio.0020329。微波处理、四氧化锇/锇膜增强膜的方法以及铜铅块状染色。标本制备程序可在http://www.stanford.edu/~redhair/JoAnn_Buchanan处获得。