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Study of SEM preparation artefacts with correlative microscopy: Cell shrinkage of adherent cells by HMDS-drying

 
: Katsen-Globa, Alisa; Puetz, Norbert; Gepp, Michael M.; Neubauer, Julia C.; Zimmermann, Heiko

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Postprint urn:nbn:de:0011-n-4048594 (863 KByte PDF)
MD5 Fingerprint: 6a77fbe8bd663a64f11e061eb7ecc4d8
This is the peer reviewed version of the following article: Study of SEM preparation artefacts with correlative microscopy: Cell shrinkage of adherent cells by HMDS-drying, which has been published in final form at 10.1002/sca.21310. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.
Created on: 01.03.2017


Scanning 38 (2016), No.6, pp.625-633
ISSN: 0161-0457
ISSN: 1932-8745
English
Journal Article, Electronic Publication
Fraunhofer IBMT ()
light microscopy (LM); scanning electron microscopy (SEM); cell shrinkage; cell spreading area; hexamethyldisalazane (HDMS)

Abstract
One of the often reported artefacts during cell preparation to scanning electron microscopy (SEM) is the shrinkage of cellular objects, that mostly occurs at a certain time-dependent stage of cell drying. Various methods of drying for SEM, such as critical point drying, freeze–drying, as well as hexamethyldisilazane (HMDS)-drying, were usually used. The latter becomes popular since it is a low cost and fast method. However, the correlation of drying duration and real shrinkage of objects was not investigated yet. In this paper, cell shrinkage at each stage of preparation for SEM was studied. We introduce a shrinkage coefficient using correlative light microscopy (LM) and SEM of the same human mesenchymal stem cells (hMSCs). The influence of HMDS-drying duration on the cell shrinkage is shown: the longer drying duration, the more shrinkage is observed. Furthermore, it was demonstrated that cell shrinkage is inversely proportional to cultivation time: the longer cultivation time, the more cell spreading area and the less cell shrinkage. Our results can be applicable for an exact SEM quantification of cell size and determination of cell spreading area in engineering of artificial cellular environments using biomaterials.

: http://publica.fraunhofer.de/documents/N-404859.html