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Noncontact versus contact imaging: An atomic force microscopic study on hepatic endothelial cells in vitro
Author(s) -
Braet Filip,
De Zanger Ronald,
Kämmer Steffan,
Wisse Eddie
Publication year - 1997
Publication title -
international journal of imaging systems and technology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.359
H-Index - 47
eISSN - 1098-1098
pISSN - 0899-9457
DOI - 10.1002/(sici)1098-1098(1997)8:2<162::aid-ima3>3.0.co;2-8
Subject(s) - biomedical engineering , scanning electron microscope , atomic force microscopy , materials science , sinusoid , resolution (logic) , biophysics , nanotechnology , composite material , pathology , biology , computer science , medicine , artificial intelligence
Liver sinusoidal endothelial cells (LEC) contain fenestrae, which control the exchange of fluids, solutes, and particles between the sinusoidal blood and the microvillous surface of the parenchymal cells. The surface of LEC can be imaged by scanning electron microscopy (SEM) and atomic force microscopy (AFM). SEM and AFM images of LEC can be used to study dynamic changes in fenestrae by comparing specimens fixed after different experimental treatments. In this article, we report the different results obtained when contact (using a constant force) or noncontact (amplitude detection) imaging on the same cells was applied. Special attention was paid on the optimalization of the image acquisition of fenestrae, because quality SEM examinations of fenestrae have already extensively been described. The following advances and conclusions are presented here: 1) High‐resolution imaging of slightly fixed LEC in fluid can be performed in noncontact AFM; 2) correct acquisition of images of fenestrae with regard to their size (Ø, ±200 nm) and shape (oval, without deformation) under liquid was possible with noncontact AFM, which was hitherto only feasible with fixed, dried, and coated LEC in contact AFM or SEM; 3) this mode of operation is more gentle to cells than contact mode; 4) images of LEC obtained in noncontact mode are of higher quality and are devoid of smearing artefacts, prominently present in contact‐mode images; 5) it is of great importance to optimize feedback and scan parameters to obtain correct surface information; and 6) LEC isolated and cultured by our method are physiologically responsive and represent an ideal object for AFM studies, because the cells are thin, smooth, and well attached to the culture substrate and show dynamic fine structural details, such as fenestrae and coated pits, which cannot be seen by light microscopy. © 1997 John Wiley & Sons, Inc. Int J Imaging Syst Technol, 8, 162–167, 1997