显微镜知识库

徕卡显微系统的知识库提供有关显微镜学科的科学研究和教学材料。内容旨在对显微镜初学者、有经验的显微镜操作实践者和使用显微镜的科学家在他们的日常工作和实验有所帮助。这里有探索交互式教程和应用笔记，你可以找到你需要的显微镜的基础知识以及前沿技术——快来加入徕卡显微知识社区，分享您的专业知识！

Serial Lift-Out workflow for cryo-ET lamella preparation - Schematic illustration of the Serial Lift-Out method applied to a vitrified biological sample (e.g., C. elegans larva) embedded in bulk vitreous ice (blue). Image courtesy: Prof. Dr. Jürgen M. Plitzko - doi.org/10.1038/s41592-023-02113-5.

Cryo-ET Sample Preparation: From Waffle Method to Serial Lift-Out

Cryo-ET sample preparation becomes more demanding when specimens are thicker, larger, or more complex. This webinar brings together four perspectives on how high-pressure freezing can be connected…

Waffle method lamella preparation and cryo-ET results - Representative workflow and results of lamella preparation from a vitrified C. elegans L1 larva using the waffle method. Image courtesy: Prof. Dr. Jürgen M. Plitzko - doi.org/10.1038/s41592-023-02113-5-

Waffle Method Workflow: From HPF to Cryo-ET Lamellae

Waffle freezing provides an HPF-based route to cryo-ET sample preparation. This workflow guide follows the process from grid and carrier assembly to vitrification, cryo-FIB milling, lamella…

3D rendering of a mouse beta cell with mitochondria (blue), insulin SGs (orange), microtubules (red), nucleus (yellow), and plasma membrane (transparent).

High-Pressure Freezing Protocols for Ultrastructural 3D EM

High pressure freezing (HPF) can help preserve hydrated cells and tissues close to their biological state at the moment of immobilization, supporting more reliable ultrastructural interpretation than…

Electron microscope (EM) image of a cross section of C. elegans (roundworm). Courtesy of T. Müller-Reichert, MPI-CBG, Dresden, Germany and K. McDonald, University of California, Berkeley, USA.

高压冷冻简介

水是细胞最主要的组成部分，因此对于维持细胞超微结构至关重要。目前，冷冻固定是固定细胞成分，而不导致其显著结构变化的唯一途径。现阶段有两种常见的方法：投入冷冻与高压冷冻固定。

"Waffle方法"：使用高压冷冻制备复杂样品

本文介绍了一种特殊的高压冷冻方法，即 "Waffle 方法 "的优点。了解 "Waffle 方法 "如何使用电镜载网作为高压冷冻的载体，从而减少样本厚度并支持复杂生物样本的高效低温电子显微镜工作流程。此外，本文还强调了现代 HPF 系统-徕卡微系统公司 EM ICE 的优势，并列举了 EM ICE 用于 "Waffle方法 "的参考文献。