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).
Original language | English |
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Pages (from-to) | 527-530 |
Number of pages | 4 |
Journal | Journal of Structural Biology |
Volume | 183 |
Issue number | 3 |
DOIs | |
Publication status | Published - Sept 2013 |
Externally published | Yes |
Keywords
- High-pressure freezing
- Adherent cells
- Cryo-sectioning
- CEMOVIS