A novel method for high-pressure freezing of adherent cells for frozen hydrated sectioning and CEMOVIS

作者：Mesman R J^*

来源：Journal of Structural Biology, 2013, 183(3): 527-530.

DOI：10.1016/j.jsb.2013.06.014

• High-pressure freezing
• Adherent cells
• Cryo-sectioning
• CEMOVIS

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).

• 出版日期2013-9