Related Experiment Video
Updated: Jun 20, 2026

Plunge Freezing: A Tool for the Ultrastructural and Immunolocalization Studies of Suspension Cells in Transmission Electron Microscopy
Published on: May 5, 2017
High pressure freezing, electron microscopy, and immuno-electron microscopy of Tetrahymena thermophila basal bodies
Janet B Meehl1, Thomas H Giddings, Mark Winey
1Department of MCD Biology, University of Colorado, Boulder, CO, USA.
Abstract:
Preservation of Tetrahymena thermophila basal body ultrastructure for visualization by transmission electron microscopy is improved by a combination of high pressure freezing (HPF) and freeze substitution (FS). These methods also reliably retain the antigenicity of cellular proteins for immuno-electron microscopy, which enables the precise localization of green fluorescent protein (GFP)-tagged and native basal body proteins. The plastic-embedded samples generated by these methods take full advantage of higher resolution visualization techniques such as electron tomography. We describe protocols for cryofixation, FS, immunolabeling, and staining. Suggestions for trouble shooting and evaluation of specimen quality are discussed. In combination with identification and manipulation of a rapidly expanding list of basal body-associated gene products, these methods are being used to increase our understanding of basal body composition, assembly, and function.
Related Concept Videos
Cryo-electron Microscopy
Transmission Electron Microscopy
Immunogold Electron Microscopy
Electron Microscope Tomography and Single-particle Reconstruction
Electron Tomography
Electron tomography can be performed either in TEM or STEM (scanning transmission...
Preparation of Samples for Electron Microscopy

