Comparative study of the conditions required to image live human epithelial and fibroblast cells using atomic force microscopy.
Successful imaging of living human cells using atomic force microscopy (AFM) is influenced by many variables including cell culture conditions, cell morphology, surface topography, scan parameters, and cantilever choice. In this study, these variables were investigated while imaging two morphologically distinct human cell lines, namely LL24 (fibroblasts) and NCI H727 (epithelial) cells. The cell types used in this study were found to require different parameter settings to produce images showing the greatest detail. In contact mode, optimal loading forces ranged between 2-2.8 x 10(-9) and 0.1-0.7 x 10(-9) (N) for LL24 and NCI H727 cells respectively. In tapping (AC) mode, images of LL24 cells were obtained using cantilevers with a spring constant of at least 0.32 N/m, while NCI H727 cells required a greater spring constant of at least 0.58 N/m. To obtain tapping mode images, cantilevers needed to be tuned to resonate at higher frequencies than their resonance frequencies to obtain images. For NCI H727 cells, contact mode imaging produced the clearest images. For LL24 cells, contact and tapping mode AFM produced images of comparable quality. Overall, this study shows that cells with different morphologies and surface topography require different scanning approaches and optimal conditions must be determined empirically to achieve images of high quality.
Murphy
Mark F
MF
General Engineering Research Institute, Liverpool John Moores University, Liverpool L3 3AF, UK. m.f.murphy@2003.ljmu.ac.uk
Lalor
Michael J
MJ
Manning
Francis C R
FC
Lilley
Francis
F
Crosby
Steven R
SR
Randall
Catherine
C
Burton
David R
DR
D002460:Cell Line; D004847:Epithelial Cells / Q000648:ultrastructure*; D005347:Fibroblasts / Q000648:ultrastructure*; D006801:Humans; D018625:Microscopy, Atomic Force / Q000379:methods*