High-Pressure Freezing and Low-Temperature Processing of Seeds for Electron Microscopy and Electron Tomography

High-pressure freezing/freeze substitution has been used to preserve biological samples for ultrastructure study instead of chemical fixation. For most plant samples, the water content is too high and cannot be properly preserved during cryofixation. Additionally, the cell wall is a barrier that pre...

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Bibliographic Details
Published inMethods in molecular biology (Clifton, N.J.) Vol. 2841; p. 207
Main Authors Li, Juan, Gou, Liangpeng, Shen, Jinbo, Cao, Wenhan
Format Journal Article
LanguageEnglish
Published United States 2024
Subjects
Arabidopsis - ultrastructure
Cold Temperature
Cryopreservation - methods
Electron Microscope Tomography - methods
Freeze Substitution - methods
Freezing
Microscopy, Electron, Transmission - methods
Pressure
Seeds - ultrastructure
Electron tomography
Freeze substitution
High-pressure freezing
Transmission electron microscopy
Secretory protein
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Summary:High-pressure freezing/freeze substitution has been used to preserve biological samples for ultrastructure study instead of chemical fixation. For most plant samples, the water content is too high and cannot be properly preserved during cryofixation. Additionally, the cell wall is a barrier that prevents the substitution of water with the resin. In this chapter, we will discuss modified high-pressure freezing and subsequent processing protocols based on our routinely used methodology for examining Arabidopsis seeds in transmission electron microscopy and electron tomography.
ISSN:1940-6029
DOI:10.1007/978-1-0716-4059-3_20