Hi everybody,
i am totally new to the programm Image J and i tried to use it to estimate my confluency in my cell culture. I found some descriptions but they do not fit for my picture at all... could anybody explain to me, what to do and how to use the programm on my cells? Please find a picture attached.
Thank you so much!
Dear Anika Dreier ,
I would recommend to set the image mode to 8 bit, then to use Process-> find Edges and then to threshold the empty (black) areas, and used the Analyse -> measure function to obtain the area fraction of the empty areas in a single fov (but of cause you would have aquired and processed a few images).
Please find a step by step process attached.
Best Soenke
Great! Thank you! that worked! Why do i set the threshold to the empty areas and not to the cells?
Hi Sönke Weinert , mega! Vielen lieben Dank für deine Hilfe!!
eine Frage hätte ich noch: Aus welchem Grund stellt man den threshold auf die freien Flächen und nicht auf die Zellen?
Es hat nach deiner Anleitung auf jeden Fall geklappt! Danke!!
Viele Grüße
Anika
My answer to the question "Why do i set the threshold to the empty areas and not to the cells?" would be, that it is simple to threshold the uniform background in stead of the quite ununiform cell layer. But that could depend on the image and cell type. If it was only round cells (i.e. in the field of hematology BaF3) the opposite might be adequate. Also, non specific fluorescent staining (CellTracker or CalceinAM) might work best with a threshold for the cells without any filter applied.
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