A multiple model probability hypothesis density tracker for time-lapse cell microscopy sequences

Abstract. Quantitative analysis of the dynamics of tiny cellular and subcellular structures in time-lapse cell microscopy sequences requires the development of a reliable multi-target tracking method capable of tracking numerous similar targets in the presence of high levels of noise, high target density, maneuvering motion patterns and intricate interactions. The linear Gaussian jump Markov system probability hypothesis density (LGJMS-PHD) filter is a recent Bayesian tracking filter that is well-suited for this task. However, the existing recursion equations for this filter do not consider a state-dependent transition probability matrix. As required in many biological applications, we propose a new closed-form recursion that incorporates this assumption and introduce a general framework for particle tracking using the proposed filter. We apply our scheme to multi-target tracking in total internal reflection fluorescence microscopy (TIRFM) sequences and evaluate the performance of our filter against the existing LGJMS-PHD and IMM-JPDA filters

A Framework for Generating Realistic Synthetic Sequences of Total Internal Reflection Fluorescence Microscopy Images

Since generation of reliable ground truth annotation of fluorescence microscopy sequences is usually a laborious and expensive task, many proposed detection and tracking methods have been evaluated using synthetic data with known ground truth. However, d

Bayesian multi-target tracking: application to total internal reflection fluorescence microscopy

This thesis focuses on the problem of automated tracking of tiny cellular and sub-cellular structures, known as particles, in the sequences acquired from total internal reflection fluorescence microscopy (TIRFM) imaging technique. Our primary biological motivation is to develop an automated system for tracking the sub-cellular structures involving exocytosis (an intracellular mechanism) which is helpful for studying the possible causes of the defects in diseases such as diabetes and obesity. However, all methods proposed in this thesis are generalized to be applicable for a wide range of particle tracking applications. A reliable multi-particle tracking method should be capable of tracking numerous similar objects in the presence of high levels of noise, high target density and complex motions and interactions. In this thesis, we choose the Bayesian filtering framework as our main approach to deal with this problem. We focus on the approaches that work based on detections. Therefore, in this thesis, we first propose a method that robustly detects the particles in the noisy TIRFM sequences with inhomogeneous and time-varying background. In order to evaluate our detection and tracking methods on the sequences with known and reliable ground truth, we also present a framework for generating realistic synthetic TIRFM data. To propose a reliable multi-particle tracking method for TIRFM sequences, we suggest a framework by combining two robust Bayesian filters, the interacting multiple model and joint probabilistic data association (IMM-JPDA) filters. The performance of our particle tracking method is compared against those of several popular and state-of-the art particle tracking approaches on both synthetic and real sequences. Although our approach performs well in tracking particles, it can be very computationally demanding for the applications with dense targets with poor detections. To propose a computationally cheap, but reliable, multi-particle tracking method, we investigate the performance of a recent multi-target Bayesian filter based on random finite theory, the probability hypothesis density (PHD) filter, on our application. To this end, we propose a general framework for tracking particles using this filter. Moreover, we assess the performance of our proposed PHD filter on both synthetic and real sequences with high level of noise and particle density. We compare its results from both aspects of accuracy and processing time against our IMM-JPDA filter. Finally, we suggest a framework for tracking particles in a challenging problem where the noise characteristic and the background intensity of sequences change during the acquisition process which make detection profile and clutter rate time-variant. To deal with this, we propose a bootstrap filter using another type of the random finite set based Bayesian filters, the cardinalized PHD (CPHD) filter, composed of an estimator and a tracker. The estimator adaptively estimates the required meta parameters for the tracker such as clutter rate and the detection probability while the tracker estimates the state of the targets. We evaluate the performance of our bootstrap on both synthetic and real sequences under these time-varying conditions. Moreover, its performance is compared against those of our other particle trackers as well as the state-of-the art particle tracking approaches

Automatic approach for spot detection in microscopy imaging based on image processing and statistical analysis

Abstract: In biological research, fluorescence microscopy has become one of the vital tools used for observation, allowing researchers to study, visualise and image the details of intracel-lular structures which result in better understanding of biology. However, analysis of large numbers of samples is often required to draw statistically verifiable conclusions. Automated methods for analysis of microscopy image data make it possible to handle large datasets, and at the same time reduce the risk of bias imposed by manual techniques in the image analysis pipeline. This work covers automated methods for extracting quan-titative measurements from microscopy images, enabling the detection of spots resulting from different experimental conditions. The work resulted in four main significant con-tributions developed around the microscopy image analysis pipeline. Firstly, an investiga-tion into the importance of spot detection within the automated image analysis pipeline is conducted. Experimental findings show that poor spot detection adversely affected the remainder of the processing pipeline...D.Ing. (Electrical and Electronic Engineering