A novel method for high-pressure freezing of adherent cells for frozen hydrated sectioning and CEMOVIS
Related Experiment Video
Updated: May 9, 2026
Electron Cryotomography of Bacterial Cells
Published on: May 6, 2010
1Membrane Enzymology, Department of Chemistry, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands; Biomolecular Imaging, Department of Biology, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands; Institute of Biomembranes, Utrecht University, Padualaan 8, 3584 CH Utrecht, The Netherlands.
Abstract:
With the development of Cryo Electron Microscopy Of Vitreous Sections (CEMOVIS), imaging cells in a close to native state has become a reality. However with the commonly used carriers for high-pressure freezing and cryo-sectioning, adherent grown cells either need to be detached from their substrate. Here a new method is presented for high-pressure freezing adherent growing cells for frozen-hydrated sectioning and CEMOVIS. Cells are cultured on golden grids, containing a carbon coated Formvar film, and frozen on a membrane carrier which provides the grids with the structural support needed to withstand the strain of trimming and cryo-sectioning. This method was successfully tested for the two different types of high-pressure freezers, those using a pressure chamber (HPM010, EMHPF, Wohlwend Compact 01/02, HPM100) and those directly pressurizing the sample (EMPact series).
More Related Videos
13:35Plunge Freezing: A Tool for the Ultrastructural and Immunolocalization Studies of Suspension Cells in Transmission Electron Microscopy
Published on: May 5, 2017
07:33Biological Sample Preparation by High-pressure Freezing, Microwave-assisted Contrast Enhancement, and Minimal Resin Embedding for Volume Imaging
Published on: March 19, 2019
Related Concept Videos
01:28Cryo-electron Microscopy
01:03Fixation and Sectioning
The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...
01:20Preparation of Samples for Electron Microscopy