Translocation Assay Example


#1

Hi, I’m a new user of Cellprofiler. This software looks great!!. Now I’m doing a project in cytoplasm-nucleus translocation assay. I try to figure out the example. But I couldn’t open the excel form of the result data (experiment.xls). It seems that the column of the data is exceeding the capacity of the excel. I need your suggestion how can i solve this problem.
Thanks in advance.


#2

Hi,
sorry for the delay. I think I have already answered your question in another post, right? If not, please give me more detail on the problem that you are having with the Experiment file.

thanks,
martha


#3

Indeed, the answer is posted here: How to open the Experiment file?

Anne


#4

Hi… I would like to ask about the Translocation measurement again.

My project is to screen the compounds in Cytoplasmic-Nuclear translocation assay. I made images in Hoechst (Blue) Channel and GFP (green) channel.
The pipeline that I used are as below :

Load Images
loadtext
Colortogray
Colortogray
Identifyprimautomatic
identifyprimautomatic
Identifysecondary
Identifysecondary
Identifytertiarysubregion(propagation)
Identifytertiarysubregion(Distance-N, 6 pixel )
Measureobjectintensity
measureobjectintensity
Calculate ratio(Mean Intensity of Distcytoplasm/Nuclei on Green image)
Classifyobjects
calculatestatictic
exporttoexcel

So, Am I right in selecting the pipeline? I couldn’t get the different ratio of green intensity in cytoplasm and nuclei in whole my samples.
I’m not sure how to determine the cytoplasmic compartment of my cells. And at the end, how can I measure the translocation events ( by using integrated intensity or mean intensity?). Shall I measure the texture and the shape of the cells?

I’m so sorry if my questions are not clear enough for you. But please help me to solve this problem.
I’m really looking forward to your answer

best,
ichwan


#5

Hi Ichwan,

It’s been a very long time since this thread was touched - apologies! Yes, the CalculateRatio module should give you the best CNT metric. Is your CNT assay working OK now?

David