Curcumin and Quercetin treatment

Hi y’all! I hope you guys all had a good break, and I’m glad you are here with me today! It was a lot of fun, but kind of stressful too, but it was definitely a new experience for me. 

This week, I ran the experiments for curcumin and quercetin individually (meaning not the combined experiment). When I went in on Monday though, I received a big shock – there were no more 96-well plates in stock at the lab, and for me to complete the experiment in time, I had to start this week. So essentially, my mentor and I decided to discuss what we could do, and we decided to go with 48 well plates instead. These wells are a bit bigger, so instead of 100 microliters of cells and 100 microliters of treatment, it changed to 200 microliters of cells and 200 microliters of treatment. Thus, the calculations for the seeding and the serial dilutions all became a bit different, but because I already created a template in excel, all I had to do was just type in the new numbers, and I got all the information I needed. 

Additionally, we created a new plate map – so instead of 3 wells with the same treatment amount, it became 2 wells with the same treatment amount (for repetition within the 1 assay). The time it takes for the experiment is essentially 5 days. Monday was Day -1, where I trypsinized the cells, and made the seeding solution for them. 

Then, I seeded them (200 microliters of the cells) into 3 48 well plates – because I will be taking measurements after 24 hours, 48 hours, and 72 hours – so I needed to seed 1 plate per time stamp. I then put them into the incubator for them to grow. 

Tuesday was Day 0, where I treated the cells with quercetin and curcumin. I calculated the amount of microliters of pure quercetin stock solution and curcumin stock solution I need to do the serial dilutions. 

I then put two hundred microliters of the treatment into the well according to the plate map design, and then I also placed a negative control (of just medium and the treatment) to make sure that the fluorescence is coming from the cells and not from the color of the chemicals (both quercetin and curcumin are yellowish-orange). 

Wednesday was Day 1 – where I measured the MFI using the plate reader Alamar Blue assay for the 24 hour seeded plate. 

Thursday was Day 2 – where I measured the MFI using the plate reader Alamar Blue assay for the 48 hour seeded plate. 

Friday was Day 2 – where I measured the MFI using the plate reader Alamar Blue assay for the 72 hour seeded plate. 

Results seem to be looking good, although I will graph them next week.