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Immunoelectron Microscopy of Cryofixed Freeze-Substituted Yeast

Fišerová, Jindřiška; Richardson, Christine; Goldberg, Martin W.


Jindřiška Fišerová

Christine Richardson


Steven D. Schwartzbach

Omar Skalli

Thomas Schikorski


Immunolabeling electron microscopy is a challenging technique with demands for perfect ultrastructural and antigen preservation. High-pressure freezing offers an excellent way to fix cellular structure. However, its use for immunolabeling has remained limited because of the low frequency of labeling due to loss of protein antigenicity or accessibility. Here we present a protocol for immunogold labeling of the yeast Saccharomyces cerevisiae that gives specific and multiple labeling while keeping the finest structural details. We use the protocol to reveal the organization of individual nuclear pore complex proteins and the position of transport factors in the yeast Saccharomyces cerevisiae in relation to actual transport events.


Fišerová, J., Richardson, C., & Goldberg, M. W. (2016). Immunoelectron Microscopy of Cryofixed Freeze-Substituted Yeast. In S. D. Schwartzbach, O. Skalli, & T. Schikorski (Eds.), High-Resolution Imaging of Cellular Proteins (243-258). Humana Press.

Online Publication Date Aug 12, 2016
Publication Date 2016-08
Deposit Date Oct 13, 2016
Publisher Humana Press
Pages 243-258
Series Title Methods in Molecular Biology
Series Number 1474
Book Title High-Resolution Imaging of Cellular Proteins
ISBN 9781493963508