Biomaterial Database

High-Pressure Freezing and Freeze Substitution for Transmission Electron Microscopy Imaging and Immunogold-Labeling. 2021

Marisa S Otegui

Department of Botany, University of Wisconsin-Madison, Madison, WI, USA. otegui@wisc.edu.

Electron microscopy enables the unbiased imaging of organelles and cellular structures at nano-meter scale resolution. The combination of cryofixation/freeze-substitution methods with other imaging techniques such as correlative light and electron microscopy (CLEM), electron tomography (ET), and immunogold-labeling provides unique opportunities to understand structural changes associated with cellular processes. This chapter presents the main steps in the preparation of Arabidopsis thaliana roots, cotyledons, anthers, and developing seeds by high-pressure freezing and freeze-substitution for structural analysis and immunogold-labeling using transmission electron microscopy.

UI	MeSH Term	Description	Entries
D011312	Pressure	A type of stress exerted uniformly in all directions. Its measure is the force exerted per unit area. (McGraw-Hill Dictionary of Scientific and Technical Terms, 6th ed)	Pressures
D013194	Staining and Labeling	The marking of biological material with a dye or other reagent for the purpose of identifying and quantitating components of tissues, cells or their extracts.	Histological Labeling,Staining,Histological Labelings,Labeling and Staining,Labeling, Histological,Labelings, Histological,Stainings
D017110	Freeze Substitution	A modification of the freeze-drying method in which the ice within the frozen tissue is replaced by alcohol or other solvent at a very low temperature.	Substitution, Freeze
D017360	Arabidopsis	A plant genus of the family BRASSICACEAE that contains ARABIDOPSIS PROTEINS and MADS DOMAIN PROTEINS. The species A. thaliana is used for experiments in classical plant genetics as well as molecular genetic studies in plant physiology, biochemistry, and development.	Arabidopsis thaliana,Cress, Mouse-ear,A. thaliana,A. thalianas,Arabidopses,Arabidopsis thalianas,Cress, Mouse ear,Cresses, Mouse-ear,Mouse-ear Cress,Mouse-ear Cresses,thaliana, A.,thaliana, Arabidopsis,thalianas, A.
D046529	Microscopy, Electron, Transmission	Electron microscopy in which the ELECTRONS or their reaction products that pass down through the specimen are imaged below the plane of the specimen.	Electron Diffraction Microscopy,Electron Microscopy, Transmission,Microscopy, Electron Diffraction,Transmission Electron Microscopy,Diffraction Microscopy, Electron,Microscopy, Transmission Electron
D055032	Electron Microscope Tomography	A tomographic technique for obtaining 3-dimensional images with transmission electron microscopy.	Electron Tomography,Tomography, Electron,Tomography, Electron Microscope,EM Tomography,Electron Microtomography,STEM Tomography,Scanning Transmission Electron Microscopy Tomography,TEM Tomography,Transmission Electron Microscopy Tomography,Transmission Electron Microtomography,Electron Microtomography, Transmission,Microscope Tomography, Electron,Microtomography, Electron,Microtomography, Transmission Electron,Tomography, EM,Tomography, STEM,Tomography, TEM