I am using Poly-L-Lysine coated 96 well plates. The cells are attached throughout the experiment. At the final step (after adding mtt), some of the formazan were coming out along with the medium while pipetting out the medium....Kindly give me suggestions..
Hi!
LNCaP cells are not an easy material unfortunately. I had similar observation while I was doing MTT assay on LNCaP cells. I also tried some coated plates or attachment factors but they didn't make thing better. I achieve the best results when I use paper towel instead of aspirating or pipetting the medium out of the wells. Maybe you can try do this in this way. After incubation with drugs you can remove the medium by inverting the plate on ethanol-soaked paper towels instead of aspirating (but very slowly and gently) and do the same after incubation with mtt mixture.
On the other hand there are also viability tests in which you can add a substrate directly to the cell culture medium and after incubation just measure the absorbance without removing growing medium at all.
Good luck!
Hi!
LNCaP cells are not an easy material unfortunately. I had similar observation while I was doing MTT assay on LNCaP cells. I also tried some coated plates or attachment factors but they didn't make thing better. I achieve the best results when I use paper towel instead of aspirating or pipetting the medium out of the wells. Maybe you can try do this in this way. After incubation with drugs you can remove the medium by inverting the plate on ethanol-soaked paper towels instead of aspirating (but very slowly and gently) and do the same after incubation with mtt mixture.
On the other hand there are also viability tests in which you can add a substrate directly to the cell culture medium and after incubation just measure the absorbance without removing growing medium at all.
Good luck!
Thank you for the suggestion, I have followed your suggestion and optimized the procedure.
Good discussion. Thanks for the update @Emilia. I am new to prostate cancer field and your reply helps a lot :D
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