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==Modifying BF Confocal Images== *Right now the BF image is mostly gray and doesn't look good (is too bright) when overlayed onto rolony images with ImageJ *Going to use GIMP to modify BF image ===Procedure=== #Open BF image in GIMP #Adjust (increase) Contrast to improve visibility of cell edges #*Can also play with brightness #Blur/Erode #Edge-detect #Thresholding #Gaussian Blur with radius 3 #Edge Detect Difference of Guardians Radius 1: 1.0 Radius 2: 16.0 #*Don't Invert #Threshold so as much black as possible without losing any cells #Brightness: -83 #Open secondary rolony polished bw images as layers #Change mode to RGB, switch black bg to alpha (transparent), and add color to each layer of rolonies #Flatten image and save <!-- #Adjust (increase) Contrast to improve visibility of cell edges #*Can also play with brightness #Blur/Erode #Edge-detect #Thresholding --first time-- #Adjust brightness and contrast #Invert #Select left and threshold #Select invert and threshold--> ===Results=== Pos1: [[File:NCIH1975_Pos1_Complete.png | 450px]]<br> Pos2: [[File:NCIH1975_Pos2_Complete.png | 450px]]<br> Pos3: [[File:NCIH1975_Pos3_Complete.png | 450px]]<br>
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