Atomic force microscopy investigation of the dependence of cellular elastic moduli on glutaraldehyde fixation |
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| Authors: | J L HUTTER† J CHEN† W K WAN†‡ S UNIYAL§ M LEABU§ & B M C CHAN§† |
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| Affiliation: | Department of Physics &Astronomy;;Biomedical Engineering Graduate Program;and; Department of Chemical &Biochemical Engineering, The University of Western Ontario, London, Ontario, Canada; Department of Microbiology and Immunology, The University of Western Ontario and the Biotherapeutics Research Group, Robarts Research Institute, London, Ontario, Canada |
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| Abstract: | The atomic force microscope (AFM) has provided nanoscale analyses of surfaces of cells that exhibit strong adhesive and cell spreading properties. However, it is frequently reported that prior fixation is required for reliable imaging of cells with lower adhesive properties. In the present study, the AFM is used to assess the effects of fixation by glutaraldehyde on the elastic modulus of a human rhabdomyosarcoma transfectant cell line RDX2C2. Our results show a sharp increase in the elastic modulus for even mild fixation (0.5% glutaraldehyde for 60 s), accompanied by a dramatic improvement in imaging reproducibility. An even larger increase is seen in NIH-3T3 mouse fibroblasts, although in that case fixation is not typically necessary for successful imaging. In addition, our results suggest that treatment with glutaraldehyde restricts the content of the resulting images to features nearer to the cell surface. |
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| Keywords: | Atomic force microscopy cell fixation cellular elastic modulus |
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