A map of sphingosine 1-phosphate distribution in the spleen

نویسندگان

  • Willy D. Ramos-Perez
  • Victoria Fang
  • Diana Escalante-Alcalde
  • Michael Cammer
  • Susan R. Schwab
چکیده

—————————————————————————————————————— macro "Reporter Regions" { /* Instructions * Draw ROI's and save as .zip file * set thresholds for each channel in the variables at the top of the macro * open the .tif image to be analyzed * follow on screen instructions to select the folders to pull the ROI.zip files and where to save the files Macro functions-Changes the background pixels in RFP and GFP channels to a value of zero-Divides GFP image by RFP image and creates a resulting image where each pixel's values are the ratio of GFP to RFP for that pixel.-Generates a membrane mask based on the CD169 and F480 stains.-Opens the pre-saved ROI files for white pulp, red pulp, and marginal zones-Gives the average GFP:RFP ratio in each ROI */ var thresholdGFP = 0; var thresholdRFP = 25; var thresholdCD169 = 20; var thresholdF480 = 32; //also try RFP at 45 for a stricter gate //10.02 is the maximum ratio if the RFP threshold is 25. //get image attributes name = getInfo("image.filename"); original = getImageID(); originalTitle = getTitle(); //Set the directory where files will be pulled from and saved to OpenDir=getDirectory("Choose the folder where data will be pulled from"); DataDir=getDirectory("Choose the folder where data will be saved"); //convert to 32 bit run("32-bit"); // split channels and threshold the GFP and RFP images. //If GFP=0, the ratio image will be 0, and will become NaN during background->NaN //If RFP=0, the ratio will be x/0, which should return NaN, bc I set Misc options

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عنوان ژورنال:

دوره 16  شماره 

صفحات  -

تاریخ انتشار 2015