Globally Optimal Cell Tracking using Integer Programming

We propose a novel approach to automatically tracking cell populations in time-lapse images. To account for cell occlusions and overlaps, we introduce a robust method that generates an over-complete set of competing detection hypotheses. We then perform detection and tracking simultaneously on these hypotheses by solving to optimality an integer program with only one type of flow variables. This eliminates the need for heuristics to handle missed detections due to occlusions and complex morphology. We demonstrate the effectiveness of our approach on a range of challenging sequences consisting of clumped cells and show that it outperforms state-of-the-art techniques.Comment: Engin T\"uretken and Xinchao Wang contributed equally to this wor

Identification of Fluorescent Compounds with Non-Specific Binding Property via High Throughput Live Cell Microscopy

10.1371/journal.pone.0028802PLoS ONE71

Detecting and Tracking Cells using Network Flow Programming

We propose a novel approach to automatically detecting and tracking cell populations in time-lapse images. Unlike earlier ones that rely on linking a predetermined and potentially under-complete set of detections, we generate an overcomplete set of competing detection hypotheses. We then perform detection and tracking simultaneously by solving an integer program to find an optimal and consistent subset. This eliminates the need for heuristics to handle missed detections due to occlusions and complex morphology. We demonstrate the effectiveness of our approach on a range of challenging image sequences consisting of clumped cells and show that it outperforms state-of-the-art techniques

Detecting cells and analyzing their behaviors in microscopy images using deep neural networks

The computer-aided analysis in the medical imaging field has attracted a lot of attention for the past decade. The goal of computer-vision based medical image analysis is to provide automated tools to relieve the burden of human experts such as radiologists and physicians. More specifically, these computer-aided methods are to help identify, classify and quantify patterns in medical images. Recent advances in machine learning, more specifically, in the way of deep learning, have made a big leap to boost the performance of various medical applications. The fundamental core of these advances is exploiting hierarchical feature representations by various deep learning models, instead of handcrafted features based on domain-specific knowledge. In the work presented in this dissertation, we are particularly interested in exploring the power of deep neural network in the Circulating Tumor Cells detection and mitosis event detection. We will introduce the Convolutional Neural Networks and the designed training methodology for Circulating Tumor Cells detection, a Hierarchical Convolutional Neural Networks model and a Two-Stream Bidirectional Long Short-Term Memory model for mitosis event detection and its stage localization in phase-contrast microscopy images”--Abstract, page iii

Comparison of NCD and CNN based mitotic classification of neural stem cells in phase contrast microscopy

Automated mitosis detection is a major difficulty in segmentation and tracking algorithms. This thesis explores the implementation of an automated mitotic detection algorithm for phase-contrast data into a segmentation and tracking platform called LEVER2. We implement two separate classification algorithms - one based on an estimate of the pairwise normalized compression distance (NCD) and one deep learning implementation using convolutional neural networks (CNN) - in combination with local statistics in order to limit the search region for mitotic events. The CNN provided a significantly higher detection rate while the NCD provided a significantly lower false positive rate. The NCD classifier generated an overall detection rate of 0.736 and an overall false positive rate of 0.0083, while the CNN approach generated an overall detection rate of 0.880 and a false positive rate of 0.0567. However, it was also determined that using a bi-modal classifier incorporating the peak normalized cross-correlation of the image with the t - 1 frame the CNN implementation could outperform the NCD classifier - achieving a false alarm rate of 0.0079 with a detection rate of 0.809 by thresholding out cell images that were above a threshold peak cross-correlation.M.S., Electrical Engineering -- Drexel University, 201

Spatio-temporal cell cycle phase analysis using level sets and fast marching methods.

Enabled by novel molecular markers, fluorescence microscopy enables the monitoring of multiple cellular functions using live cell assays. Automated image analysis is necessary to monitor such model systems in a high-throughput and high-content environment. Here, we demonstrate the ability to simultaneously track cell cycle phase and cell motion at the single cell level. Using a recently introduced cell cycle marker, we present a set of image analysis tools for automated cell phase analysis of live cells over extended time periods. Our model-based approach enables the characterization of the four phases of the cell cycle G1, S, G2, and M, which enables the study of the effect of inhibitor compounds that are designed to block the replication of cancerous cells in any of the phases. We approach the tracking problem as a spatio-temporal volume segmentation task, where the 2D slices are stacked into a volume with time as the z dimension. The segmentation of the G2 and S phases is accomplished using level sets, and we designed a model-based shape/size constraint to control the evolution of the level set. Our main contribution is the design of a speed function coupled with a fast marching path planning approach for tracking cells across the G1 phase based on the appearance change of the nuclei. The viability of our approach is demonstrated by presenting quantitative results on both controls and cases in which cells are treated with a cell cycle inhibitor

Nanoparticles in medicine: Automating the analysis process of high-throughput microscopy data.

Automated tracking of cells across timelapse microscopy image sequences typically employs complex segmentation routines and/or bio-staining of the tracking objective. Often accurate identification of a cell's morphology is not of interest and the accurate segmentation of cells in pursuit of non-morphological parameters is complex and time consuming. This thesis explores the potential of internalized quantum dot nanoparticles as alternative, bio- and photo-stable optical markers for tracking the motions of cells through time. CdTe/ZnS core-shell quantum dots act as nodes in moving light display networks within A549, epithelial, lung cancer cells over a 40 hour time period. These quantum dot fluorescence sources are identified and interpreted using simplistic algorithms to find consistent, non-subjective centroids that represent cell centre locations. The presented tracking protocols yield an approximate 91% success rate over 24 hours and 78% over the full 40 hours. The nanoparticle moving light displays also provide simultaneous collection of cell motility data, resolution of mitotic traversal dynamics and identification of familial relationships enabling the construction of multi-parameter lineage trees. This principle is then developed further through inclusion of 3 different coloured quantum dots to create cell specific colour barcodes and reduce the number of time points necessary to successfully track cells through time. The tracking software and identification of parameters without detailed morphological knowledge is also demonstrated through automated extraction of DOX accumulation profiles and Cobalt agglomeration accruement statistics from two separate toxicology assays without the need for cell segmentation