Hello,
I will start an experiment using macrophage (THP-1 treated with PMA). I have ordered the plate that suitable for fluorescence plate reader which is glass 24 wells black uncoated tissue culture plate. My question regarding culturing macrophage, should I coat them with poly L lysine for culturing THP-1 differentiated macrophage? As we know that macrophages are very sticky if I put them in the uncoated plate is the behaviour of cells changed? (not adhere on glass)
If I coat the plate and seed the cells on them, treat the cells with nanosensor, technically (using plate reader), Can I take the reading of fluorescence intensity from the bottom or top of the plate?
Thanks
Reem
Hi,
In principle, white tissue culture plate reflects light and the black ones quench the light. Are you planning any chemiluminescence studies? I would suppose to coat the already differentiated macrophages with poly L lysine and avoid using any uncoated one. Black wells could be used for chemiluminescence studies or florescence ones.
Culture of macrophage (differentiated THP1) dont need coating in general tissue culture grade plates. However, I never used black tissue culture plates. Coating with poly L lysine will not have an effect unless you are expecting shape changes or crawling.
If the plate is of tissue culture grade, there is no need for you to coat the plate with poly L lysine. PMA treated THP-1 cells stick well and show well differentiated morphology. At the same time , there is no wrong even if you prefer to do the coating.
Regarding your 2nd query about reading the fluorescence intensity, it depends upon the kind of plate reader and the plate. Is the plate with solid walls and clear bottoms?
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