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For quantifying cell speed and motility, 3D coordinates of cells in each frame were recorded and calculated using Volocity.
At each time point we fit a fourth order polynomial through the coordinates of cells in this boundary region.
They segmented given tissue images using a k-means algorithm and generated different cell graphs using the positional coordinates of cells for each segmented image.
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The average expression levels of these genes (y-axis in Fig. 2: cell lineage index) can thus be used as surrogates for the coordinates of cell's positions on each trajectory (x-axis in Fig. 2).
To generate smoothed texture coding maps of the normalized barrel each cell was assigned set of point coordinates c k (x i, y i ) with i = 1…N k, N k —number of cells found for texture k, and c k —color assigned to a texture k and x i, y i are the spatial coordinates of cell i.
We have established Drosophila male germline stem cells (GSCs) as a model to study the checkpoint that coordinates polarization of cells and cell division (Cheng et al., 2008; Inaba et al., 2010; Yuan et al., 2012).
This microscope system captures images of dyed cell nuclei, computes the location coordinates of individual cells, and synchronizes this with the voice coil motor stage and single-particle irradiation system consisting of a scintillation counter and a beam deflector.
Three-dimensional coordinates of the cells were analyzed statistically to determine whether those cells activated by movements of the arm and leg were segregated spatially.
For each experimental condition we evaluated the fluorescent emission from (typically) ~200 individual cells by first determining the physical locations (pixel coordinates) of single cells in a phase contrast image.
To quantify the emission levels of individual cells in the bioluminescence images, we first used the dark field images to obtain the pixel coordinates of individual cells that had remained immobile during the experiment.
In subsequent imaging sessions, the coordinates of these cells combined with overview images at low (5×) magnification are then used to retrieve the position of defined cells (for details, see methods).
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