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. 2011 Mar;10(1):9-15.
doi: 10.1109/TNB.2011.2113397. Epub 2011 Feb 24.

Quantitative analysis of human keratinocyte cell elasticity using atomic force microscopy (AFM)

Carmen Kar Man Fung  1 , Ning Xi, Ruiguo Yang, Kristina Seiffert-Sinha, King Wai Chiu Lai, Animesh A Sinha
Affiliations

Quantitative analysis of human keratinocyte cell elasticity using atomic force microscopy (AFM)

Carmen Kar Man Fung et al. IEEE Trans Nanobioscience. 2011 Mar.

Abstract

We present the use of atomic force microscopy (AFM) to visualize and quantify the dynamics of epithelial cell junction interactions under physiological and pathophysiological conditions at the nanoscale. Desmosomal junctions are critical cellular adhesion components within epithelial tissues and blistering skin diseases such as Pemphigus are the result in the disruption of these components. However, these structures are complex and mechanically inhomogeneous, making them difficult to study. The mechanisms of autoantibody mediated keratinocyte disassembly remain largely unknown. Here, we have used AFM technology to image and measure the mechanical properties of living skin epithelial cells in culture. We demonstrate that force measurement data can distinguish cells cultured with and without autoantibody treatment. Our demonstration of the use of AFM for in situ imaging and elasticity measurements at the local, or tissue level opens potential new avenues for the investigation of disease mechanisms and monitoring of therapeutic strategies in blistering skin diseases.

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Figures

Fig. 1
Fig. 1
Typical deflection-displacement curves obtained for glass and cell surface. Glass is chosen as the hard material compared with the cell (soft material) and the slope of the curve for the glass is 1. δ is the indentation depth of the cell.
Fig. 2
Fig. 2
A typical force-indentation curve of the cell surface.
Fig. 3
Fig. 3
Immunofluoresence image of the fixed HaCaT cells. The arrows indicate desmoplakin (stained for FTIC), one of the main proteins in the desmosomal protein family, distributed in the peripheral of the cell. This confirms that the cell forms desmosome between their neighbors. The AFM images were taken at those boundaries to visualize the detailed structure of the cell-cell adhesion structure.
Fig. 4
Fig. 4
A typical AFM image of living HaCaTs. The cell shape and overall structure between two cells are clearly shown in a height image [scan size: 20 × 20 µm2]. The arrows indicate the cell-cell junction between two adjacent keratinocytes.
Fig. 5
Fig. 5
Force measurements of living HaCaT cells. (a) The numbered marks indicate the positions where force–displacement curve measurements were obtained (overlaid on a height image to visualize HaCaT cell topography). (b) Deflection–displacement curves measured on the living HaCaT cell at positions marked in Fig. 4(a). The arrow indicates the contact point of the cell and the AFM tip.
Fig. 6
Fig. 6
Comparison of deflection-displacement curves of living HaCaT cells from the same sample with (A) goat anti-mouse Ig irrelevant control antibody and (B) 1h post pathogenic anti-Dsg3 antibody treatment.
Fig. 7
Fig. 7
Comparison of force-indentation curves of living HaCaT cells from the same sample with (A) goat anti-mouse Ig irrelevant control antibody and (B) 1h post pathogenic anti-Dsg3 antibody treatment.
Fig. 8
Fig. 8
Hertzian fitting of force-indentation curves of living HaCaT cells from the same sample with (A) goat anti-mouse Ig irrelevant control antibody and (B) 1h post pathogenic anti-Dsg3 antibody treatment. For the control sample (P=12.01154), the E then can be calculated as 84.5KPa, while for the pathogenic antibody treated sample (P=31.25158), the Young’s modulus would be 247 KPa. The Young’s modulus increased three-fold after antibody treatment. Red line indicates the fitting using the Hertz model, and black indicates the experimental data.
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