Spindle Frap-Flip Analyzer
Analyze protein dynamics within spindle
The purpose of this macro is to follow the intensity dynamics occurring within double labeled microtubule spindle. Intensity changes that are occurring at stable position is easy, but this macro enables measurement of intensity changes that accompanies movement of the bleached/activated position.
Using a pair of two-channel image stacks, this macro measures dynamics of integrated intensity profile along x-axis of a channel in a masked region created using the other channel.
ImageJ ver 1.34j or higher
Download K_YprojectionV2.ijm to a place where you can access easily. Install the macro by [Plugins→Macros→Install…].
Let’s see what it does using a two-channel image data. In red channel (Fig. 1), tubulin is labeled and in the green channel (Fig. 3), certain microtubule-associated protein is labeled. The program first creates a mask (black and white image, Fig. 2) from tubulin R-channel that specifies the area of the spindle. Then this specified area is measured for the integrated intensity profile (projected in y-axis, and measured along x-axis). Same procedure is done for the all frames within the stacks automatically. Profile data will be printed out in the Results window as a table. Another function included in the macro will let the user to plot the multiple profiles in a graph to evaluate the measurements. Results could also be exported and examined using other software such as Excel.
Fig.1 Red Channlel/Tubulin.