The idea is to draw a line from the center of the cell that passes through the bleb. The simplest way to do so is to use kymographs. We would like to get the speed at which the front of the bleb grows. Bleb growth dynamicsįirst, get the raw transmitted movie here (22MB). Goal of this study is to get the dynamics of the bleb growth and retraction. After a while, a new cortex is polymerized and the bleb retracts. This leads to the formation of a bleb, a woughly spherical bulge of membrane, initially devoid of cortex. When ablated at the left spot, the constraint on volume is relaxed, and the cortex can now contracts, expulsing some cytoplasm through the hole, like a relaxing ball that would be squeezed by your hands. The ablation laser power saturates the transmitted detector, which results in white frames in the movie. At rest, this tension gives the floating cell this spherical shape.Ī picosecond pulsed laser allowed us to make a micro ablation of the cortex, without disrupting the membrane. The acto-myosin cortex is the part of the cytoskeletton that lies just below the membrane This cortex is active and generate a contractile force that presses on the cell volume. The transmitted signal is overlayed with a confocal image of the GFP channel in various ways. This is a movie made in the lab of Ewa Paluch, MPI-CBG, featuring a floating L929 cell (mouse fibroblast), stained for myosin (RLC-GFP) being ablated in the cortex. We will introduce them in a real-world example, using the most simple functions in ImageJ. So we get an image where we move along space in the x direction and along time in the y direction.Įxamples of usage can be found in the references listed at the end of this page. These lines are stacked along the y axis for all frames. Each time point gives a intensity line, plotted e.g. They can be seen as a x-t scan, where the intensity along a given line is plotted for all images of a stack. They are particularly useful to monitor in a glimpse a moving organelle and characterize its motion. Kymographs are a way to represent on a single image a dynamic process. If you’d like to help, check out the how to help guide! Introduction The content of this page has not been vetted since shifting away from MediaWiki.
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