Hi,
I am enquiring if anyone knows any microscope platform that allows for live-display of normalised fluorescence intensity of field of view?
For example if a GFP reporter that can be normalised to a co-transfected RFP internal control.
As the microscope has to switch channels I envision that there are two ways of doing this:
1) With a time-lag, the microscope captures both fluorescent channels and displays the normalised pixel intensity.
2) Real-time display using dual colour capture microscope and displaying the normalised pixel intensity
I've tried searching online but have not found anything useful.
Why I want to do this is to pick up cells under the microscope that respond to our GFP reporter.
Hi !
1. Cheap option: get micro-manager (https://micro-manager.org/), write your own script to make the live calculations of the ratio (GFP/mCherry).
2. Commercial option: Visitron is selling it (http://www.visitron.de/Products/Software/VisiView_Ratio_Option/visiview_ratio_option.html)
Good luck !
Hi !
1. Cheap option: get micro-manager (https://micro-manager.org/), write your own script to make the live calculations of the ratio (GFP/mCherry).
2. Commercial option: Visitron is selling it (http://www.visitron.de/Products/Software/VisiView_Ratio_Option/visiview_ratio_option.html)
Good luck !
Ah yes exactly that, ratiometric measurement. I just never knew a good name to search google with. We have Metamorph for olympus microscope and doesn't have this function so we have to do the processing using imageJ after capturing the two separate channels. Do you know of other software that offer this?
Another poster suggested micro-manager, which is free for writing code.
We don't have a FRET setup so I guess it will always a slight lag time but is useable for colony selection.
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