I am planning to use a 96-well format for an experiment as this will greatly improve throughput and expedite treatments. I will be using Caco-2 cells cultured apically. I need them to form confluent, differentiated monolayer on the insert. Do you have any experience doing this in a 96-well format? What are the difficulties you have encountered? Thanks!
I used 96-wells for bacterial biofilm experiments, however the same difficulties apply. Medium overflow and cross contamination of treatments (if any used), too little media and fast dryness, loss of tracking treatments if a spreadsheet or 96-well map wasn't properly made, contaminating control wells by sample wells media by bumping or tilting the plate by mistake, fast growing cells need to be checked every 24hrs not to risk death by apoptosis, seeding cells must be titrated according to cells' division capacity/rate as not overgrow the wells, Nucleaic acids extraction in limited quantities. A major advantage though is multiple testing of different treatments and ability to test effect of different treatments on expression in a comparative setting minimizing human/environmental variables. Hope this helps.
Rabeah Al-Temaimi thank you for your response! Yes, I don't plan to use the 96 well format for nucleic acid analysis. Will probably use it for viability assays and others. Might also adjust my media replenishment from every 48H to every day. Again, thanks!
I've tried doing something similar (transwell 96-well). The edge effect was the biggest problem. Another issue I can foresee in your experiment is testing the permeability of each of the monolayer (96). If the integrity of some of them isn't good then you would have to avoid those wells and it might make the assay design confusing. Anyway, good luck.
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