PRO OR CONS LOCAL VS. GLOBAL IMAGERY INFORMATION FOR IDENTIFYING CELL MIGRATORY POTENTIAL (original) (raw)
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Informatik aktuell, 2013
Methods to evaluate migration capacity of stem cells and the inhibition by chemicals are important for biomedical research. Here, we established an automated image processing framework to quantify migration of human neural crest (NC) cells into an initially empty, circular region of interest (ROI). The ROI is partially filled during the experiment by migrating cells. Based on an image captured only once at the end of the biological experiment, the framework identifies the initial ROI. The identification worked also, when the distribution of surrounding cells showed large heterogeneity. After segmentation, the number of migrated cells was identified. The image processing framework was capable of efficiently quantifying chemical effects on cell migration.
A novel approach to characterizing cell migration
2019
Cancer cell metastasis has been responsible for the vast majority of cancer-related deaths in the United States. The processes involved in cancer cell metastasis, such as extravasation and intravasation, are driven by cell motility. The conventional method of characterizing cell motility typically involved imaging live cells under a microscope for several hours and tracking cells’ trajectories manually with the help of computer software, and this method is highly inefficient and time-consuming for obtaining cell motility information. This dissertation aims to develop a new method to quantitatively characterize cell motility based on the spatial distribution of cells in clones at a specific time point. A simulation study was first performed to evaluate the correlation between cell spatial distribution in the clones and cell motility. Clonal distributions of cells were generated at the 72 hr time point from computer-simulated cell trajectories based on the PRW model for clone sizes of...
A simple method to study cellular migration
Journal of Neuroscience Methods, 2004
We describe here a simple and fast method for the characterisation of cell motion. By projecting on a single plane different positions of the cell a ribbon is generated, whose characteristics can be related to the type of motion. The proposed method allows both to determine, very quickly, the motility of a population of cells and to investigate and characterise properties of a single cell’s motion. The methodology presented here can be applied to a large range of cell movement and also adapted and extended to other problems involving biological motion.
Taking Aim at Moving Targets in Computational Cell Migration
Trends in Cell Biology, 2015
Cell migration is central to the development and maintenance of multicellular organisms. Fundamental understanding of cell migration can, for example, direct novel therapeutic strategies to control invasive tumor cells. However, the study of cell migration yields an overabundance of experimental data that require demanding processing and analysis for results extraction. Computational methods and tools have therefore become essential in the quantification and modeling of cell migration data. We review computational approaches for the key tasks in the quantification of in vitro cell migration: image pre-processing, motion estimation and feature extraction. Moreover, we summarize the current state-of-the-art for in silico modeling of cell migration. Finally, we provide a list of available software tools for cell migration to assist researchers in choosing the most appropriate solution for their needs. Computational Cell Migration in a Nutshell Cell migration plays a fundamental role in physiological phenomena including neural development, wound healing, and immune function, as well as in disorders such as neurological diseases, fibrosis, and cancer metastasis [1-8]. Investigation of cell migration is therefore essential for successful intervention in physiological and pathological phenomena [9-12]. A major driver in the advance of cell migration research has been the evolution of instrumentation (microscopes and cameras) and the corresponding development of experimental tools and biological models. Indeed, 2D in vitro assays [13,14] have recently given way to more sophisticated two-and-a-half-dimensional (2.5D) and 3D approaches [15,16] which more faithfully represent the tissue environment. Because in vivo experiments are difficult and costly, in vitro and ex vivo setups are widely used, especially in drug compound and gene screening [17,18]. This review therefore primarily focuses on the quantification of cell migration in in vitro setups, while we refer the reader to specific literature on in vivo work [19-23].
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Journal of Microscopy, 2012
Many fundamental biological processes, such as the search for food, immunological responses and wound healing, depend on cell migration. Video microscopy allows the magnitude and direction of cell migration to be documented. Here, we present a simple and inexpensive method for simultaneous tracking of hundreds of migrating cells over periods of several days. Low-magnification dark-field microscopy was used to visualize individual cells whereas time-lapse video images were acquired by computer for future analysis. We employed an automated tracking algorithm to identify individual cells on each video image allowing migration paths to be tracked using a nearest neighbour algorithm. To test the method, we followed the time-course of migration of 3T3 fibroblasts, endothelial cells and individual amoeba in the absence of any chemical stimulus gradient. All cell types showed a 'random walk' behaviour in which mean squared displacement in position increased linearly with time. We defined a 'migration coefficient' (D mig ), analogous to a diffusion coefficient, which gave an estimate of cell migration rate. D mig depended on cell type and temperature. When amoebas were made to undergo chemotaxis, the cells no longer followed a random walk but instead moved at a near constant velocity (V av ) towards the chemotactic stimulus.
2012 9th IEEE International Symposium on Biomedical Imaging (ISBI), 2012
The understanding of the embryogenesis in living systems requires reliable quantitative analysis of the cell migration throughout all the stages of development. This is a major challenge of the "in-toto" reconstruction based on different modalities of "in-vivo" imaging techniques-spatio-temporal resolution and image artifacts and noise. Several methods for cell tracking are available, but expensive manual interaction-time and human resources-is always required to enforce coherence. Because of this limitation it is necessary to restrict the experiments or assume an uncontrolled error rate. Is it possible to obtain automated reliable measurements of migration? can we provide a seed for biologists to complete cell lineages efficiently? We propose a filtering technique that considers trajectories as spatio-temporal connected structures that prunes out those that might introduce noise and false positives by using multi-dimensional morphological operators.
Automated time-lapse microscopy and high-resolution tracking of cell migration
Cytotechnology, 2006
We describe a novel fully automated high-throughput time-lapse microscopy system and evaluate its performance for precisely tracking the motility of several glioma and osteoblastic cell lines. Use of this system revealed cell motility behavior not discernable with conventional techniques by collecting data (1) from closely spaced time points (minutes), (2) over long periods (hours to days), (3) from multiple areas of interest, (4) in parallel under several different experimental conditions. Quantitation of true individual and average cell velocity and path length was obtained with high spatial and temporal resolution in “scratch” or “wound healing” assays. This revealed unique motility dynamics of drug-treated and adhesion molecule-transfected cells and, thus, this is a considerable improvement over current methods of measurement and analysis. Several fluorescent vital labeling methods commonly used for end-point analyses (GFP expression, DiO lipophilic dye, and Qtracker nanocrystal...
An Image-based Dynamic High-throughput Analysis of Adherent Cell Migration
Bio-protocol, 2021
In this protocol, we describe a method to monitor cell migration by live-cell imaging of adherent cells. Scratching assay is a common method to investigate cell migration or wound healing capacity. However, achieving homogenous scratching, finding the optimal time window for end-point analysis and performing an objective image analysis imply, even for practiced and adept experimenters, a high chance for variability and limited reproducibility. Therefore, our protocol implemented the assessment for cell mobility by using homogenous wound making, sequential imaging and automated image analysis. Cells were cultured in 96-well plates, and after attachment, homogeneous linear scratches were made using the IncuCyte ® WoundMaker. The treatments were added directly to wells and images were captured every 2 hours automatically. Thereafter, the images were processed by defining a scratching mask and a cell confluence mask using a software algorithm. Data analysis was performed using the IncuC...
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2017
We describe an integrated experimental-computational pipeline for quantifying cell migration in vitro. This pipeline is robust to image noise, open source, and user friendly. The experimental component uses the Oris cell migration assay (Platypus Technologies) to create migration regions. The computational component of the pipeline creates masks in Matlab (MathWorks) to cell-covered regions, uses a genetic algorithm to automatically select the migration region, and outputs a metric to quantify the migration of cells. In this work we demonstrate the utility of our pipeline by quantifying the effects of a drug (Taxol) and of the secreted Anterior Gradient 2 (sAGR2) protein in the migration of MDA-MB-231 cells (a breast cancer cell line). In particular, we show that blocking sAGR2 reduces migration of MDA-MB-231 cells.