Count and calculate positive stained Cells Immunofluorescence Staining in ratio to stained nuclei

Image Analysis
#1

hey everyone.

I’m trying to create a pipeline to count and calculate the percentage of positive stained cells (intracellular) in ratio to all stained nuclei (DAPI) in a immunofluorescence staining experiment.
I already figured out to identify respective objects. now I’m struggeling with the modules and settings to count those objects and the calculation.
I used IDENTIFY_PRIMARY_OBJECTS, which does a good job.

Can anyone please help me? Which modules to I have to add with which settings?

Thanks a lot

Flo

#2

Hi @Flo,

This is my understanding,

  1. DAPI - Total number of Nuclei (i.e. total number of Cells)
  2. +ive stain - Number of +ive stain cells
    If you have already identified nuclei & +ive cells then use,
    “MeasureObjectShapeSize” to count the number & use “CalculateMath” module to find your ratio

Hope this helps.

Regards,
Lakshmi
www.wakoautomation.com