Drying cells for SEM, AFM and TEM by hexamethyldisilazane: a study on hepatic endothelial cells.
Critical point drying (CPD) is a common method of drying biological specimens for scanning electron microscopy (SEM). Drying by evaporation of hexamethyldisilazane (HMDS) has been described as a good alternative. This method, however, is infrequently used. Therefore, we reassessed HMDS drying. Cultured rat hepatic sinusoidal endothelial cells (LEC), possessing fragile fenestrae and sieve plates, were subjected to CPD and HMDS drying and evaluated in the scanning electron microscope, atomic force microscope (AFM) and transmission electron microscope (TEM). We observed no differences between the two methods regarding cellular ultrastructure. In contrast with CPD, HMDS drying takes only a few minutes, less effort, low costs for chemicals and requires no equipment. We conclude that HMDS-dried specimens have equal quality to CPD ones. Furthermore, the method also proved useful for drying whole-mount cells for TEM and AFM.
Braet
F
F
Laboratory for Cell Biology and Histology, Free University of Brussels, Belgium.
De Zanger
R
R
Wisse
E
E
D000818:Animals; D002478:Cells, Cultured; D004727:Endothelium / Q000648:ultrastructure; D005069:Evaluation Studies as Topic; D005404:Fixatives; D016591:Histocytological Preparation Techniques; D008099:Liver / Q000648:ultrastructure*; D018625:Microscopy, Atomic Force / Q000379:methods; D008854:Microscopy, Electron / Q000379:methods; D008855:Microscopy, Electron, Scanning / Q000379:methods; D017646:Organosilicon Compounds; D051381:Rats
D005404:Fixatives; D017646:Organosilicon Compounds; C024548:hexamethylsilazane