Methods for Cell and Particle Tracking

Biomedical Imaging Group Rotterdam, Departments of Medical Informatics and Radiology, Erasmus MC-University Medical Center Rotterdam, Rotterdam, The Netherlands.
Methods in enzymology (Impact Factor: 2.09). 12/2012; 504:183-200. DOI: 10.1016/B978-0-12-391857-4.00009-4
Source: PubMed


Achieving complete understanding of any living thing inevitably requires thorough analysis of both its anatomic and dynamic properties. Live-cell imaging experiments carried out to this end often produce massive amounts of time-lapse image data containing far more information than can be digested by a human observer. Computerized image analysis offers the potential to take full advantage of available data in an efficient and reproducible manner. A recurring task in many experiments is the tracking of large numbers of cells or particles and the analysis of their (morpho)dynamic behavior. In the past decade, many methods have been developed for this purpose, and software tools based on these are increasingly becoming available. Here, we survey the latest developments in this area and discuss the various computational approaches, software tools, and quantitative measures for tracking and motion analysis of cells and particles in time-lapse microscopy images.

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    • "Six image series of bulge stage and mature root hair, and 24 of growing root hairs were used to define the endosomal population. The speed and distances in individual, interacting, and clustered endosomes were measured in growing root hairs using the semiautomatic MtrackJ function in ImageJ (Meijering et al., 2012). vonWangenheim et al.Endosomes were selected and speed, distance, and intensity were determined as parameters during structure displacement. "
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    ABSTRACT: The dynamic localization of endosomal compartments labeled with targeted fluorescent protein tags is routinely followed by time lapse fluorescence microscopy approaches and single particle tracking algorithms. In this way trajectories of individual endosomes can be mapped and linked to physiological processes as cell growth. However, other aspects of dynamic behavior including endosomal interactions are difficult to follow in this manner. Therefore we characterized the localization and dynamic properties of early and late endosomes throughout the entire course of root hair formation by means of spinning disc time lapse imaging and post-acquisition automated multitracking and quantitative analysis. Our results show differential motile behavior of early and late endosomes and interactions of late endosomes that may be specified to particular root hair domains. Detailed data analysis revealed a particular transient interaction between late endosomes – termed herein as dancing-endosomes – which is not concluding to vesicular fusion. Endosomes preferentially located in the root hair tip interacted as dancing-endosomes and traveled short distances during this interaction. Finally, sizes of early and late endosomes were addressed by means of super-resolution structured illumination microscopy (SIM) to corroborate measurements on the spinning disc. This is a first study providing quantitative microscopic data on dynamic spatio-temporal interactions of endosomes during root hair tip growth.
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    • "To differentiate between synaptic boutons and trafficking synaptic vesicles, tracks were created using a nearest-neighbour linking approach for objects moving less than 500nm within 3 frames. Tracks were visualized and quantified with the MTrackJ plugin of Fiji[20].The identification of different unloading profiles, the weak and strong unloading synaptic boutons, was performed as described before[21]. In R statistical programming language (version 3.1.1[15]), "

    Full-text · Article · Dec 2015 · BMC Neuroscience
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    • "Left epifluorescence picture; right total internal reflection fluorescence (TIRF) micrograph of the basal cell area (Color figure online) E. Merklinger et al.: No Evidence for Spontaneous Lipid Transfer… occurred. The movement of spots was manually tracked using the MTrackJ plugin (Meijering et al. 2012) for ImageJ applying centroid positioning within ROIs of 7 or 9 pixel diameter. Mean-square displacement (MSD) was calculated as described previously (Qian et al. 1991; Kusumi et al. 1993) for all spots within a 50 9 50 pixel ROI which were traceable for C20 frames before and after treatment. "
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    ABSTRACT: Non-vesicular lipid transport steps play a crucial role in lipid trafficking and potentially include spontaneous exchange. Since membrane contact facilitates this lipid transfer, it is most likely to occur at membrane contact sites (MCS). However, to date it is unknown whether closely attached biological membranes exchange lipids spontaneously. We have set up a system for studying the exchange of lipids at MCS formed between the endoplasmic reticulum (ER) and the plasma membrane. Contact sites were stably anchored and the lipids cholesterol and phosphatidylcholine (PC) were not capable of transferring spontaneously into the opposed bilayer. We conclude that physical contact between two associated biological membranes is not sufficient for transfer of the lipids PC and cholesterol.
    Full-text · Article · Oct 2015 · Journal of Membrane Biology
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