Hi every one....
We are intending to do some comparative migration/invasion assays in different cell lines- HCT 116, Sw480, HT29 and Colo205.
I couldn't find the cell size, so I wanted to know that if somebody has any idea how to choose a perfect pore size for a cell type and if my cells have variable diameter, do I need to use separate pore size for each one ?
One thing I would again mention, we are intended to compare the migratory potential of the se cells with one another, so to me coosing pore size is the main point and it has to be perfect so as to show a comparative result.
Any help would be highly appreciated....
Thanking you,
Regards
I know these cell lines very well these colorectal cancer cell lines show variable differences in cell size.
It will be very helpful for you to subculture on coverslips and stain with H&E stain. Then Examine under light microscope high power 40 and take images for each cell line.
Using the free Image J software you can measure the surface area and measure size of each cell line.
Then you can select the suitable pore size for each cell line.
Image J software..
Image J/ Fiji.
steps:
1. File > open > select image (open image)
2. Image > type > 8-bit (change to B & W)
3. Process > subtract background > rolling ball radius
4. Image > adjust > threshold (B & W)> apply
5. Analyse > analyse particles a. Area b. Perimeter c. Circularity d. circumference e. Feret's diameter
6. Exclude on edge. show outline, show clear results, display results
7. Transfer to excel
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