I'm having phase contrast image data of my previous cell culture where I have to count number of differentiated Mesenchymal Stem cells among normal cell population. can anyone please suggest me how to count accurately using phase contrast image .I want to calculate percentage differentiation by counting normal cell vs differentiated cells. Is it can be possible using ImageJ software?
If you have a distinct morphology difference between differentiated versus non differentiated this would be relatively simple. I would open the image in Microsoft paint. Then using the marker tool I would add a small red dot to the cells that are differentiated. You can use a different color, like blue to mark all of the ones non-differentiated. You could either manually count the number of red vs blue dots or you could enter the image into ImageJ and quantitate there. I think if you can visually not distinguish between differentiated versus undifferentiated you have no choice but to use some sort of staining or qPCR.
Determining differentiation from morphology alone is messy and depending on your cell type can be very unreliable. It is much better to have a specific protein stained that is either present or absent depending on differentiation status and then quantitate based on that.
Thank you Julie for your kind suggestion I think I can use your idea in counting cells from some of the images where cells are really not too crowded.
please read this article
Cell tracking algorithm using the phase contrast microscopy image
Published in:
SICE Annual Conference (SICE), 2012 Proceedings of
Date of Conference:
20-23 Aug. 2012
Page(s):
1926 - 1929
ISSN :
pending
Print ISBN:
978-1-4673-2259-1
INSPEC Accession Number:
13056038
Conference Location :
Akita
Publisher:
IEE
Hello Avinash,
If you know which are the differentiated cells among the normal cells in your images then the answer is yes. It can be measured easily and effectively by Image-J in cell counter mode. Infact it gives an automatic label as per your selection to all your cells so it becomes an easy exercise. With that tool, you can even count cells in 90-100% confluent confluent as all you need is just to do a mouse click and cell will be automatically taken into consideration. You can also save these images for your fiuture records and counted data can be transported easily to excell which is really easy and hassel free.
As per my view. Image J is the best option in this condition.
Hope it helps !
Ajay Thank you for the your kind suggestion I have already got the same solution before , now my problem is solved.
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