Story

Cryo-Electron Microscopy of Vitreous Sections (CEMOVIS) of Yeast

Application Note for Leica EM HPM100 - Life Science Research

Protocol
The sections are of yeast frozen with a Leica HPM100 High Pressure Freezer in the copper tube system, the cell paste was mixed with a pH 6.5 MES/dextran buffer so that a final MES concentration of 50mM and a dextran concentration of 20% was achieved.

The samples were sectioned on a Leica EM UC7/ EM FC7 Ultramicrotome with micromanipulator at -140°C and the section thickness was set to 50nm. The sections were attached to Agar lacey grids.

Authors

Topics & Tags

The sections were imaged using a Tecnai Polara 300KeV (FEI, The Netherlands) microscope fitted with a 4K Gatan CCD camera. The magnification for the sections was 23K, with a defocus of -6um for the tomogram and -8um for the projection image, and the diffraction was done with a camera length of 930mm. The image in panel A is an average of the central 10 slices of a reconstruction done with the IMOD package (Kremer et. al., 1996), image processing software from a tomogram collected using the FEI software.

Interested to know more?

Talk to our experts. We are happy to answer all your questions and concerns.

Contact Us

Do you prefer personal consulting?

  • Leica Microsystems Inc.
    1700 Leider Lane
    Buffalo Grove, IL 60089 United States
    Office Phone : +1 800 248 0123
    Service Phone : 1 800 248 0223
    Fax : +1 847-236-3009

You will find a more detailed list of local contacts here.