Abstract

Computer-based fully-automated cell tracking is becoming increasingly important in cell biology, since it provides unrivalled capacity and efficiency for the analysis of large datasets. However, automatic cell tracking’s lack of superior pattern recognition and error-handling capability compared to its human manual tracking counterpart inspired decades-long research. Enormous efforts have been made in developing advanced cell tracking packages and software algorithms. Typical research in this field focuses on dealing with existing data and finding a best solution. Here, we investigate a novel approach where the quality of data acquisition could help improve the accuracy of cell tracking algorithms and vice-versa. Generally speaking, when tracking cell movement, the more frequent the images are taken, the more accurate cells are tracked and, yet, issues such as damage to cells due to light intensity, overheating in equipment, as well as the size of the data prevent a constant data streaming. Hence, a trade-off between the frequency at which data images are collected and the accuracy of the cell tracking algorithms needs to be studied. In this paper, we look at the effects of different choices of the time step interval (i.e., the frequency of data acquisition) within the microscope to our existing cell tracking algorithms. We generate several experimental data sets where the true outcomes are known (i.e., the direction of cell migration) by either using an effective chemoattractant or employing no-chemoattractant. We specify a relatively short time step interval (i.e., 30 s) between pictures that are taken at the data generational stage, so that, later on, we may choose some portion of the images to produce datasets with different time step intervals, such as 1 min, 2 min, and so on. We evaluate the accuracy of our cell tracking algorithms to illustrate the effects of these different time step intervals. We establish that there exist certain relationships between the tracking accuracy and the time step interval associated with experimental microscope data acquisition. We perform fully-automatic adaptive cell tracking on multiple datasets, to identify optimal time step intervals for data acquisition, while at the same time demonstrating the performance of the computer cell tracking algorithms.

Highlights

  • Cell migration is a fundamental process that is essential to life and it is linked to many important physiological and pathological events such as immune response, wound healing, tissue differentiation, embryogenesis, and tumour invasion [1,2,3,4,5,6,7]

  • We evaluate the accuracy of our existing cell tracking algorithms to illustrate the effects of these different time step intervals

  • Our fully-automated approach differs from the current-state-of-the-art cell tracking methodologies in that we investigate the dynamic interplay between segmentation and tracking and the variable adaptive time step interval for generating quality image datasets from the microscopes

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Summary

Introduction

Cell migration is a fundamental process that is essential to life and it is linked to many important physiological and pathological events such as immune response, wound healing, tissue differentiation, embryogenesis, and tumour invasion [1,2,3,4,5,6,7]. Modern microscopy with image capturing devices is at the heart of techniques to study live-cell migration, such as those driven by chemotactic signalling during directed migration [8,9,10,11]. The advent of high-throughput, high-resolution microscopy, and imaging techniques means that cell outlines and migration trajectories or pathways may be visually accessible and manually traceable by a human operator (through manual tracking) to result in what is known as the gold-standard ground truth. With the ever-increasing use of microscopy, a humongous amount of experimental data is routinely generated in many laboratories. Most of these data are collected in the absence of a monitoring segmentation and tracking algorithm. A noticeable downside of fully-automated tracking is that the recognition capability of computers is very limited (comparing to a trained human operator) despite the sizeable efforts made from the wider academic community [16,17,18,19,20]

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