Automated tracking in live-cell time-lapse movies

Youssef, Simon; Gude, Sebastian; Rädler, Joachim O.

doi:10.1039/c1ib00035g

Cited by 23 publications

(19 citation statements)

References 18 publications

(22 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Images were binarized by setting a global threshold. Binarized images were clustered and tracked with Cell Evaluator . Coordinates of the center of mass and the area of the nuclei were extracted and analyzed in MATLAB.…”

Section: Methodsmentioning

confidence: 99%

Cell Motility on Polyethylene Glycol Block Copolymers Correlates to Fibronectin Surface Adsorption

et al. 2014

Self Cite

View full text Add to dashboard Cite

Adhesion and motility of cells on polyethylene glycol (PEG) engineered surfaces are of fundamental interest for the development of biotechnological devices. Here, the structure of PEG block copolymers physisorbed to surfaces by polyLlysine (PLL) or polypropylene oxide (PPO) is studied. Cell behavior on such surfaces incubated with fibronectin (FN) is analyzed via time-lapse microscopy, the amount and the location of FN is determined via neutron reflectivity. While FN does not adsorb onto PPOPEG, 0.4-0.7 mg m(-2) of FN is found in the vicinity of the PLL moiety of PLLPEG. Cells exhibit 21% increased motility on PLLPEG (5 kDa PEG chains) compared to pure FN layers, and 12% decreased motility for PLLPEG (2 kDa PEG chains). These findings suggest that by design of PEGylated surfaces cell migration can be controlled.

show abstract

Section: Methodsmentioning

confidence: 99%

Cell Motility on Polyethylene Glycol Block Copolymers Correlates to Fibronectin Surface Adsorption

et al. 2014

Self Cite

View full text Add to dashboard Cite

show abstract

“…Binarized images were clustered and tracked with a cell evaluator. 29 Coordinates of the centre of mass and the area of nuclei were extracted and analysed in MATLAB. The position of the micro-structured lattice was evaluated from the bright-eld images for every marker position.…”

Section: Image and Data Analysismentioning

confidence: 99%

Cellular self-organization on micro-structured surfaces

2014

Self Cite

View full text Add to dashboard Cite

Micro-patterned surfaces are frequently used in high-throughput single-cell studies, as they allow one to image isolated cells in defined geometries. Commonly, cells are seeded in excess onto the entire chip, and non-adherent cells are removed from the unpatterned sectors by rinsing. Here, we report on the phenomenon of cellular self-organization, which allows for autonomous positioning of cells on micropatterned surfaces over time. We prepared substrates with a regular lattice of protein-coated adhesion sites surrounded by PLL-g-PEG passivated areas, and studied the time course of cell ordering. After seeding, cells randomly migrate over the passivated surface until they find and permanently attach to adhesion sites. Efficient cellular self-organization was observed for three commonly used cell lines (HuH7, A549, and MDA-MB-436), with occupancy levels typically reaching 40-60% after 3-5 h. The time required for sorting was found to increase with increasing distance between adhesion sites, and is well described by the time-to-capture in a random-search model. Our approach thus paves the way for automated filling of cell arrays, enabling high-throughput single-cell analysis of cell samples without losses.

show abstract

“…To analyze the time-lapse movies, we used the ImageJ plugin Cell Evaluator [Youssef, 2011]. The droplets were segmented and tracked using images recorded in brightfield mode.…”

Section: Analysis Of Time-lapse Moviesmentioning

confidence: 99%