I am using alamar blue to test the fluroescene and absorbance on two colorectal cancer cells lines.
Research/ literature I have read give different advice regarding how to seed the wells in the plates; they differ between instructing not to seed int he outer wells, and some suggest not to seed any wells next to each other.
I have 24 variables to analyse for each cell line and filling one plate for each cell line offers me the opportunity to have all variables plus controls, but how will this change my results.
If anyone has performed these and can give some feedback, this would be great
Hi, Samuel.
I have no experience of alamar blue dye, however, if you measure fluorescence by spectrofluorometry, you've got to use black-walled 96-well plate for cell culture. We used this for determine DCF-DA flurorescence for ROS estimation. If you use plain 96-well culture plate, the machine will be interrupted by reading the surrounding wells' fluorescence, too. The specific plate has only clear bottom to black-out the neighboring wells, leading to precise measurement.
Have a look at the link. ViewPlate-96 Black, Optically Clear Bottom, Tissue Culture Treated, Sterile, 96-Well with Lid (http://www.perkinelmer.com/product/viewplate-96-f-tc-50x1b-6005182). You can use all 96-wells if you use it.
Regards.
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