title: Shrinkage of freeze-dried cryosections of cells: Investigations by EFTEM and cryo-CLEM.
abstract: Freeze-drying of cryosections of cells or tissues is considered to be the most efficient preparation for microanalysis purpose related to transmission electron microscopy. It allows the measurements of ions and water contents at the ultrastructural level. However an important drawback is associated to freeze-drying: the shrinkage of the cryosections. The aim of this paper is the investigation of this phenomenon by means of three different methods applied to both hydrated and dehydrated cryosections: direct distance measurements on fiducial points, thickness measurements by energy filtered transmission microscopy (EFTEM) and cryo-correlative light electron microscopy (cryo-CLEM). Measurements in our experimental conditions reveal a lateral shrinkage around 10% but the most important result concerns the lack of differential shrinkage between most of the cellular compartments.
authors.affiliation: Laboratoire de Recherche en Nanosciences, EA4682, Université de Reims Champagne Ardenne, 21 rue Clément Ader, 51685 Reims Cedex 2, France.
authors.forename: G
authors.initials: G
authors.lastname: Casanova
authors.affiliation: Laboratoire de Recherche en Nanosciences, EA4682, Université de Reims Champagne Ardenne, 21 rue Clément Ader, 51685 Reims Cedex 2, France.
authors.forename: F
authors.initials: F
authors.lastname: Nolin
authors.affiliation: Laboratoire de Recherche en Nanosciences, EA4682, Université de Reims Champagne Ardenne, 21 rue Clément Ader, 51685 Reims Cedex 2, France.
authors.forename: L
authors.initials: L
authors.lastname: Wortham
authors.affiliation: CNRS UMR 7369, Université de Reims Champagne Ardenne, Reims, France.
authors.forename: D
authors.initials: D
authors.lastname: Ploton
authors.affiliation: Laboratoire de Recherche en Nanosciences, EA4682, Université de Reims Champagne Ardenne, 21 rue Clément Ader, 51685 Reims Cedex 2, France.
authors.forename: V
authors.initials: V
authors.lastname: Banchet
authors.affiliation: Laboratoire de Recherche en Nanosciences, EA4682, Université de Reims Champagne Ardenne, 21 rue Clément Ader, 51685 Reims Cedex 2, France. Electronic address: jean.michel@univ-reims.fr.
authors.forename: J
authors.initials: J
authors.lastname: Michel
mesh_terms: D020285:Cryoelectron Microscopy / Q000379:methods; D016616:Cryoultramicrotomy / Q000379:methods*; D004577:Electron Probe Microanalysis / Q000379:methods; D005612:Freeze Drying; D006367:HeLa Cells; D006801:Humans; D008855:Microscopy, Electron, Scanning; D061848:Optical Imaging / Q000379:methods
keywords: Cells cryosections; Cryo-CLEM; EFTEM; Freeze-drying; Shrinkage