Setting minimum and maximum Intensity + Add colours


#1

Hello.
I have a set of images (~200) similar to the attached one. Each Image has two channels. The first Channel is the GFP and the second channel is the corresponding RFP. Would that be possible for me to get some assistance to execute the following

  1. Enhance intensity.= Set the minimum and maximum intensity values for each channel as follows

                  a. GFP channel = (minimum = 10, maximum = 150) 
                      RFP channel = (minimum = 20, maximum = 450) 
    

2 Add green to GFP channel and red to RFP channel

Regards,.
Dinindu


#2

@dinindu.senanayake Here is a pipeline, forumpipeline.cppipe (8.0 KB), that should get you started. The trick to using the RescaleIntensity module is to know how the images are being normalized as they are being processed by CellProfiler. The metadata of the image you shared indicates the image will automatically be normalized between [0,1] by dividing the pixel integers by the image bit-depth, i.e. 2^16. Therefore, to get the ranges you mention in your post, divide the integer values by 2^16 and update the parameters in the RescaleIntensity module accordingly.

Here is a sample of the output.


#3

Thank you very much. This pipeline worked perfectly form my analysis