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The positioning of adhesion molecules relative to the microtopography of the cell surface has a significant influence on the molecule's availability to form adhesive contacts. Measurements of the ratio of fluorescence intensity per unit area in epi-fluorescence images versus total internal reflection fluorescence images provides a means to assess the relative accessibility for bond formation of different fluorescently labeled molecules in cells pressed against a flat substrate. Measurements of the four principal adhesion molecules on human neutrophils reveal that L-selectin has the highest ratio of total internal reflection fluorescence/epi intensity, and that P-selectin glycoprotein ligand-1 (PSGL-1) and the integrins alphaLbeta2 (LFA-1) and alphaMbeta2 (Mac-1) have ratios similar to each other but lower than for L-selectin. All of the ratios increased with increasing impingement, indicating an alteration of surface topography with increasing surface compression. These results are consistent with model predictions for molecules concentrated near the tips of microvilli in the case of L-selectin, and sequestered away from the microvillus tips in the case of LFA-1, Mac-1, and PSGL-1. The results confirm differences among adhesion molecules in their surface distribution and reveal how the availability of specific adhesion molecules is altered by mechanical compression of the surface in live cells.

The recruitment of cells to the vascular wall in vivo or the capture of cell subpopulations at the surface of a fabricated device requires the formation of bonds between specific molecular pairs on the cell and the substrate. The ability of a molecule to form a bond depends critically on its localization relative to the cell surface topography. In this report, we present a framework for the quantitative assessment of molecular availability that accounts for the deformability of the cell surface and the balance of forces in the interface, as well as the variability of surface protrusion lengths and the preference for molecules to reside at or away from the tips of surface projections. We also examined how molecular availability should change with increasing compression of the cell against the substrate. Finally, we convolved the distribution of molecules at the interface with a decaying evanescent excitation to predict the fluorescence intensity in total internal reflectance fluorescence microscopy, which can provide a quantitative measure of the relative availability of different molecules at a cell-substrate interface. Model predictions show good agreement with measurements of fluorescence intensity of different molecules labeled fluorescently on the surface of a human neutrophil compressed against a glass surface.

This article presents a stochastic model designed to analyze experimental data on the development of cell clones composed of two (or more) distinct types of cells. The proposed model is an extension of the traditional multi-type Bellman-Harris branching stochastic process allowing for nonidentical time-to-transformation distributions defined for different cell types. A simulated pseudo likelihood method has been developed for the parametric statistical inference from experimental data on cell clones under the proposed model. The method uses simulation-based approximations of the means and the variance-covariance matrices of cell counts. The proposed estimator for the vector of unknown parameters is strongly consistent and asymptotically normal under mild regularity conditions, while its variance-covariance matrix is estimated by the parametric bootstrap. A Monte Carlo Wald test is proposed for the test of hypotheses. Finite sample properties of the estimator have been studied by computer simulations. The model and associated methods of parametric inference have been applied to the analysis of proliferation and differentiation of cultured O-2A progenitor cells that play a key role in the development of the central nervous system. It follows from this analysis that the time to division of the progenitor cell and the time to its differentiation (into an oligodendrocyte) are not identically distributed. This biological finding suggests that a molecular event determining the type of cell transformation is more likely to occur at the start rather than at the end of the mitotic cycle.