Fluorescence microscopy imaging has become one of the essential tools used by biologists to visualize and study intracellular particles within a cell. Studying these particles is a long-term research effort in the field of microscopy image analysis, consisting of discovering the relationship between the dynamics of particles and their functions. However, biologists are faced with challenges such as the counting and tracking of these intracellular particles. To overcome the issues faced by biologists, tools which can extract the location and motion of these particles are essential. One of the most important steps in these analyses is to accurately detect particle positions in an image, termed spot detection. The detection of spots in microscopy imaging is seen as a critical step for further quantitative analysis. However, the evaluation of these microscopic images is mainly conducted manually, with automated methods becoming popular. This work presents some advances in fluorescence microscopy image analysis, focusing on the detection methods needed for quantifying the location of these spots. We review several existing detection methods in microscopy imaging, along with existing synthetic benchmark datasets and evaluation metrics.
Tracking of multiple bright particles (spots) in fluorescence microscopy image sequences is seen as a crucial step in understanding complex information in the cell. However, fluorescence microscopy generates high a volume of noisy image data that cannot be analysed efficiently by means of manual analysis. In this study we compare the performance of two computer-based tracking methods for tracking of bright particles in fluorescence microscopy image sequences. The methods under comparison are, Interacting Multiple Model filter and Feature Point Tracking. The performance of the methods is validated using synthetic but realistic image sequences and real images. The results from experiments show that the Interacting Multiple Model filter performed best, under the test conditions.
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