Variation in cell-substratum adhesion in relation to cell cycle phases

R. Geoff Richards, Iolo A. ap Gwynn, D. O. Meredith, G. R. Owen

Research output: Contribution to journalArticlepeer-review

24 Citations (SciVal)

Abstract

The quantification of focal adhesion sites offers an assessable method of measuring cell–substrate adhesion. Such measurement can be hindered by intra-sample variation that may be cell cycle derived. A combination of autoradiography and immunolabelling techniques, for scanning electron microscopy (SEM), were utilised simultaneously to identify both S-phase cells and their focal adhesion sites. Electron-energy ‘sectioning’ of the sample, by varying the accelerating voltage of the electron beam, combined with backscattered electron (BSE) imaging, allowed for S-phase cell identification in one energy ‘plane’ image and quantitation of immunogold label in another. As a result, it was possible simultaneously to identify S-phase cells and their immunogold-labelled focal adhesions sites on the same cell. The focal adhesion densities were calculated both for identified S-phase cells and the remaining non-S-phase cells present. The results indicated that the cell cycle phase was a significant factor in determining the density of focal adhesions, with non-S-phase cells showing a larger adhesion density than S-phase cells. Focal adhesion morphology was also seen to correspond to cell cycle phase; with ‘dot’ adhesions being more prevalent on smaller non-S-phase and the mature ‘dash’ type on larger S-phase cells. This study demonstrated that when quantitation of focal adhesion sites is required, it is necessary to consider the influence of cell cycle phases on any data collected.
Original languageEnglish
Pages (from-to)58-67
Number of pages10
JournalExperimental Cell Research
Volume293
Issue number1
DOIs
Publication statusPublished - 01 Feb 2004

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