I want to compare the cytotoxicites of a doxorubicin-only treatment and a doxorubicin-loaded magnetosome (or some other nanoparticle) treatment. Are there particular reactions between doxorubicin and tetrazolium dye which would make this method problematic? Is the MTT assay the right assay to use?
MTT assay is better than SRB assay and on the other hand a cheaper assay than LDH assays. The LDH assay kits are overpriced always.
MTT assay is a very good technique, if the cell seeding is done properly on well plates. MTT assay provides an overall quantitative cellular viability. Therefore, the cell seeding on well plates should be uniform.
For standard deviation take 3 wells per sample or condition. Also use negative and positive control for comparison.
Hi I fully support Rob....U need to confirm the outcome of MTT assay by other methods. Like trypan blue. Actually I used to do MTT assay for the neuroprotection assay but many times got faulty result, I will suggest to use Trypan blue method for your experiment, its better cheaper although cumbersome method...
I prefer to use Trypan blue assay, it's easy and very convinent. MTT assay or Crystal blue assay it's better for alive cells.
I think there's no problem if you use MTT assay for doxorubicin cytotoxic assay. I prefer recommending you the WST1 assay as I had udes it before. It's quicker and more simple than MTT is
I hyave ran MTT and MTS assays many times using doxorubicin singly and in combination with a variety of chemotherapeutic drugs. I repeat each assay twice and have triplicate columns for each treatment on each plate. I have found that this technique is very accurate, as long as all your parameters are strictly adhered to.
i think MTT or WST assay are good methods incase of cytotoxicity assays ,but if you are intrested in cardiotoxicity ,you may quantify the amount of doxorubicin in heart tissues in case oh animal expemint .
I agree with Rodi. If you have cardiac tissues from mice/rat that have been treated/Controls it may be wise to 'back-up' your assays with immunohistochemistry. I have just set up an experiment with 50 mice with controls, pre-treatments, Doxorubicin singly and in combination etc and will be harvesting the heart tissues (amongst others) for immunohistochemistry to look for cardiotoxicity/damage.
Dear Luke Lazatin,
I suggest you to also perform some oxidative stress analysis, since doxorubicin is a classic inducer of ROS damage.
Good luck
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