Mixture-model classification in DNA content analysis

Abstract

DNA abundance provides important information about cell physiology and proliferation activity. In a typical in vitro cellular assay, the distribution of the DNA content within a sample is comprised of cell debris, G0/G1-, S-, and G2/M-phase cells. In some circumstances, there may be a collection of cells that contain more than two copies of DNA. The primary focus of DNA content analysis is to deconvolute the overlapping mixtures of the cellular components, and subsequently to investigate whether a given treatment has perturbed the mixing proportions of the sample components. We propose a restricted mixture model that is parameterized to incorporate the available biological information. A likelihood ratio (LR) test is developed to test for changes in the mixing proportions between two cell populations. The proposed mixture model is applied to both simulated and real experimental data. The model fitting is compared with unrestricted models; the statistical inference on proportion change is compared between the proposed LR test and the Kolmogorov–Smirnov test, which is frequently used to test for differences in DNA content distribution. The proposed mixture model outperforms the existing approaches in the estimation of the mixing proportions and gives biologically interpretable results; the proposed LR test demonstrates improved sensitivity and specificity for detecting changes in the mixing proportions. © 2007 International Society for Analytical Cytology