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

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).

journal_name

J Struct Biol

journal_title

Journal of structural biology

authors

Mesman RJ

doi

10.1016/j.jsb.2013.06.014

subject

Has Abstract

pub_date

2013-09-01 00:00:00

pages

527-530

issue

3

eissn

1047-8477

issn

1095-8657

pii

S1047-8477(13)00179-2

journal_volume

183

pub_type

杂志文章

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