Hi! Has anyone followed the growth of cells attached to Cytodex 3 microcarriers using Presto Blue assay? Could you please provide me a detailed protocol?
I've had some difficulties with standardized the protocol, It appears that Microcarriers quench the fluorescence of the cells. (Cells without microcarriers have a higher fluorescence than cells at the same concentration with microcarriers).
Thank you in advance
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Hi Anamaria, i have the same problem, do you try in absorbance test?
Hi Alida! Good to know that I'm not the only one with the same problem. What do you mean with try the absorbance test? Do you have any protocol that you can share?
Until now, counting nuclei with violet crystal is the only way to count the cells in the microcarriers.
Hi Anamaria, the prestoblue can we test in the way of fluorescence or absorbance, I do not have the protocol with microcarriers, only with cells growing in plates, and is in the datasheet of the product https://www.thermofisher.com/document-connect/document-connect.html?url=https%3A%2F%2Fassets.thermofisher.com%2FTFS-Assets%2FLSG%2Fmanuals%2FMAN0018370-PrestoBlueCellViabilityReagent-PI.pdf&title=UHJvZHVjdCBTaGVldDogUHJlc3RvQmx1ZSBDZWxsIFZpYWJpbGl0eSBSZWFnZW50.
I have problems with the reaction because the protocol indicates that in 2 hours I can read the absorbance o fluorescence, but is not working, because at this time I do not have the change of the values between the sample and the negative control., Only in 24 hours, I can read, but I do not if it is replicable. Do you try using another test type, for example, Calcein AM?
On the other hand, do you harvest the cells growing in the citodex 3, in an easy way? because I used citodex 1, and I tried several times with different approaches, and is not working. Could you send me a suggestion of protocol?
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