Topics in living cell miultiphoton laser scanning microscopy (MPLSM) image analysis

Abstract

Multiphoton laser scanning microscopy (MPLSM) is an advanced fluorescence imaging technology which can produce a less noisy microscope image and minimize the damage in living tissue. The MPLSM image in this research is the dehydroergosterol (DHE, a fluorescent sterol which closely mimics those of cholesterol in lipoproteins and membranes) on living cell's plasma membrane area. The objective is to use a statistical image analysis method to describe how cholesterol is distributed on a living cell's membrane. Statistical image analysis methods applied in this research include image segmentation/classification and spatial analysis. In image segmentation analysis, we design a supervised learning method by using smoothing technique with rank statistics. This approach is especially useful in a situation where we have only very limited information of classes we want to segment. We also apply unsupervised leaning methods on the image data. In image data spatial analysis, we explore the spatial correlation of segmented data by a Monte Carlo test. Our research shows that the distributions of DHE exhibit a spatially aggregated pattern. We fit two aggregated point pattern models, an area-interaction process model and a Poisson cluster process model, to the data. For the area interaction process model, we design algorithms for maximum pseudo-likelihood estimator and Monte Carlo maximum likelihood estimator under lattice data setting. For the Poisson Cluster process parameter estimation, the method for implicit statistical model parameter estimate is used. A group of simulation studies shows that the Monte Carlo maximum estimation method produces consistent parameter estimates. The goodness-of-fit tests show that we cannot reject both models. We propose to use the area interaction process model in further research.