Shrinkage of freeze-dried cryosections of cells: Investigations by EFTEM and cryo-CLEM

• Published in: Micron (Oxford, England : 1993) England : 1993), 2016-09, Vol.88, p.77-83
• Main Authors: Casanova, G., Nolin, F., Wortham, L., Ploton, D., Banchet, V., Michel, J.
• Format: Article
• Language: English
• Subjects: Cells cryosections; Cryo-CLEM; Cryoelectron Microscopy - methods; Cryoultramicrotomy - methods; EFTEM; Electron Probe Microanalysis - methods; Freeze Drying; HeLa Cells; Humans; Microscopy, Electron, Scanning; Optical Imaging - methods; Shrinkage
• ISSN: 0968-4328; 1878-4291
• DOI: 10.1016/j.micron.2016.06.005

•Complete study about the phenomenon of shrinkage in freeze–dried cells cryosections.•Interest and validity of freeze-drying for TEM sample preparation when analytical measurements on cells are concerned.•First investigation of shrinkage phenomenon by cryo-CLEM including fluorescence of freeze-dried cryosections of cells. 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.