Hydra Plasma FIB DualBeam for High-resolution Cryo Auto Slice & View and Reliable Lamella Preparation for Cellular and Tissue Samples

The Hydra Plasma FIB DualBeam system is a powerful tool for cellular structure biology, providing unique capabilities for sample preparation and high-resolution imaging. Advanced features including the cryo stage, multi-ion species plasma FIB and Elstar SEM column make it ideal for imaging cellular and tissue samples. As a key application of this microscope, the cryo Auto Slice & View technique enables automated serial milling followed by imaging on vitrified biological samples. With optimized imaging parameters for the Elstar SEM, volumetric datasets with maximized contrast are achievable. In this study, we will demonstrate the capabilities of the Hydra Plasma FIB DualBeam system for cryo Auto Slice & View by showcasing whole cell volumes of Chlamydomonas reinhardtii and Chinese hamster kidney cells milled using argon ions. Additionally, we will share results of milling with different ion species on these samples. Furthermore, results from high-pressure frozen mice brain tissue showing lamella preparation and cryo Auto Slice & View will be presented. High pressure freezing is a critical technique for preserving the native state of tissues and cellular samples and the Hydra PFIB has advantages for efficient preparation of these sample types for cryo-electron microscopy. Overall, the Hydra PFIB system provides a complete solution for cellular structure biology, enabling efficient lamella preparation and high-resolution imaging with the versatility for handling a wide range of cryogenic samples. Single slice image of Chlamydomonas reinhardtii from an automated cryo slice and view data collection (1.2 kV, 13 pA, 5 nm pixel). Segmentation of 3D volume from slice and view data of Chlamydomonas reinhardtii. Whole cell segmentation was performed by Jessica Heebner (Penns State College of Medicine).