A novel method for high-pressure freezing of adherent cells for frozen hydrated sectioning and CEMOVIS.
J Struct Biol
; 183(3): 527-530, 2013 Sep.
Article
em En
| MEDLINE
| ID: mdl-23831450
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).
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Base de dados:
MEDLINE
Assunto principal:
Criopreservação
/
Microscopia Crioeletrônica
Limite:
Humans
Idioma:
En
Ano de publicação:
2013
Tipo de documento:
Article