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Foraminifera (Ammonia confertitesta) labeled with membrane-permeable calcein, high-pressure frozen in salt water using EM ICE. The sample was cryo-planed and targeted with the M205 on the Cryo-Fluo Enuity, then transferred under cryo conditions to the Cryo-Stellaris for widefield and confocal imaging, revealing details of the staining pattern. Image courtesy: David Evans, University of Southampton.

High-Pressure Freezing for Organoids: Cryo CLEM & FIB Lift Out

Master cryo EM workflow steps for challenging 3D samples: when to choose HPF vs. plunge freezing, reproducible blotting/ice control, contamination aware transfers, Cryo CLEM 3D targeting in organoids,…

Complete camera overview of EM grid recorded with 3 channels. Inserts displaying the positions, where superresolved 3D confocal images were recorded. 3D renderings of these positions are shown in the zoomed inserts. Fluorescence channels (nuclei by Hoechst, blue; mitochondria by MitoTracker Green, green; lipid Droplets by Bodipy and Crimson Beads, red). Width of a grid square is 90 ?m, width of a grid bar is 35 ?m. Samples kindly provided by Ievgeniia Zagoriy, Mahamid-Group, EMBL Heidelberg, Germany.

从显微镜到电镜：完整的冷冻光电联用工作流程

在题为“多模态玻璃化征程，从实验台到电子显微镜的冷冻关联工作流程”的网络研讨会上，专家团队（Edoardo D'Imprima、Zhengyi Yang、Andreia Pinto 和 Martin…

Correlation of markers in the LM and the FIB image.

如何对荧光结构三维定位以进行冷冻FIB切片

冷冻ET（电子断层扫描）是一种专用的透射电子显微镜技术，可以重建观察区域的三维体积。借助先进的冷冻EM（电子显微镜），图像分辨率可以提升到令人难以置信的亚纳米等级。因此，可以在细胞内的原生环境中研究蛋白质以及其他生物分子，从而揭示尚未探明的分子机制。由于细胞和组织必须薄到能够透过电子，样品必须进行切片以获取足够薄的样品体积（薄层）。为对样品中的靶区进行精确的三维定位，冷冻共聚焦显微镜是必不可少的工…

Root-hypocotyl junction of Arabidopsis thaliana. Image acquired with TauContrast. Sample courtesy: Dr. Melanie Krebs, COS, University of Heidelberg.

TauContrast 对图像复杂样本的好处

在这次访谈中，Timo Zimmermann博士谈到了他在应用TauSense工具方面的经验，以及这些工具在研究厚样本或超大胚胎等复杂样本中的潜力。作为巴塞罗那基因组调控中心（CRG）先进光学显微镜单位的负责人，Timo Zimmermann博士在2020年测试了STELLARIS 5共聚焦系统。