High Pressure Freezing
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Video Tutorials: Filling and Assembling of Different Carriers for High-Pressure Freezing
High pressure freezing (HPF) is a cryo-fixation method primarily for biological samples, but also for a variety of non-biological materials. It is a technique that yields optimal preservation in many…Read article
Electron Tomography of Cryo-Immobilized Plant Tissue: A Novel Approach to Studying 3D Macromolecular Architecture of Mature Plant Cell Walls In Situ
Cost-effective production of lignocellulosic biofuel requires efficient breakdown of cell walls present in plant biomass to retrieve the wall polysaccharides for fermentation. In-depth knowledge of…Read article
Brief Introduction to Freeze Fracture and Etching
Freeze fracture describes the technique of breaking a frozen specimen to reveal internal structures. Freeze etching is the sublimation of surface ice under vacuum to reveal details of the fractured…Read article
Capturing Cellular Dynamics with Millisecond Temporal Resolution
The combination of two powerful techniques: optogenetics and high-pressure freezing now makes it possible to visualize a dynamic cellular activity with temporal resolution of 5 milliseconds. By…Read article
Freeze Substitution of Trypanosoma brucei
Chemical fixation of biological specimens for ultrastructural investigation is a relatively slow and selective process, and therefore a common source of artifacts. Freezing, on the other hand, is an…Read article
Brief Introduction to Freeze Substitution
Freeze-substitution is a process of dehydration, performed at temperatures low enough to avoid the formation of ice crystals and to circumvent the damaging effects observed after ambient-temperature…Read article
Thermodynamic Considerations Regarding the LN2 in a High Pressure Freezer
Employing liquid nitrogen (LN2) as a coolant in the complex process of high pressure freezing raises certain considerations regarding phase transition not only of the liquid sample to be frozen but…Read article
From Dynamic Live Cell Imaging to 3D Ultrastructure: Novel Integrated Methods for High Pressure Freezing and Correlative Light-Electron Microscopy
To correlate dynamic events in adherent cells with both ultrastructural and 3D information, we developed a method for cultured cells that combines confocal time-lapse images of GFP-tagged proteins…Read article
Safe High Pressure Freezing of Infectious Microorganisms
Especially in core EM-facilities one is confronted with material (microorganisms and cells) which are infectious. It is a must to follow the safety rules according to the National regulations. However…Read article
A Precise and Rapid Mapping Protocol for Correlative Light and Electron Microscopy of small invertebrate organisms
CLEM (correlative live cell and electronmicroscopy) seeks to bridge the data acquired with different imaging strategies, typically between light microscopy and electron microscopy. It has been…Read article
Brief Introduction to High-Pressure Freezing
Water is the most abundant cellular constituent and therefore important for preserving cellular ultra-structure. Currently the only way to fix cellular constituents without introducing significant…Read article
Targeting of Peroxisomal Matrix Proteins in the Diatom Phaeodactylum Tricornutum
P. tricornutum cells expressing different types of GFP fusion proteins were harvested via centrifugation at 1,500xg and cryoimmobilized by high-pressure freezing on gold carriers.Read article
Dry Ultrathin Sectioning Combined With High Pressure Freezing
We have used cultured UMR106-01 osteoblastic cells to investigate the process of bone mineralization. UMR106-01 cells as well as primary calvarial bone cells assembly spherical extracellular…Read article
Postlipolytic Insulin-dependent Remodeling of Micro Lipid Droplets in Adipocytes
Despite the lipolysis–lipogenesis cycle being a fundamental process in adipocyte biology, very little is known about the morphological changes that occur during this process. The remodeling of lipid…Read article
CLEM: Combining the Strengths of Light and Electron Microscopy
In recent years light microscopy studies have been dominated by live cell imaging while electron microscopy has been used for high-resolution studies. Latterly, there has been increasing interest in…Read article