Yvonne Zhou- Week 4- Lewis Lab

Greetings!

I fell sick last weekend so I was not able to write the post. Below is my recollection.
I continued my work on the Mesenchymal stem cell(MSC) differentiation assay--preparing media, feeding the cells and taking pictures of them at times. More cells popped off the bottom so eventually we decided to discard the non-confluent group. It was frustrating but "that often happens when you are dealing with cells", as my mentor said. Culturing cells was somehow more difficult than caring for a pet... We considered starting over but then we decided to wait until the cells in the confluent group could be stained.
Here are the microscopy images of the cells on day 13:

Cells fed with media containing Adipogenic cocktail

Cells fed with DMEM Complete (the normal media). As many cells have aggregated together, the cells on this image have a much lower density. Basically they look similar to the cells in the Adipo group.

Cells fed with media containing Osteogenic cocktail. The dark areas in the image are the minerals that the cells create as they reach certain point in the differentiation phase.

We were to do staining the following week (which we did on Wednesday this week).
I also had more experience on the 3D printer! I wanted to be more familiar with the printers, so I decided to print a heart for fun. This is the Robomama. (Robocaster, also called Robopapa, had an accident and was down infinitely:( The lab people were to send it away for repair, which would cost a ton because of its humongous size.)

I printed on Robomama this time.

I prepared an 10:1 SE 1700 ink and added the catalyst ahead of time to help cross-linking and curing.

Trial and Error. Please excuse the simple 2D structure that I created, as it was too difficult for me, a beginner at coding, to design a fun 3D structure on a fused deposition printer. 

My code worked differently from what I would expect it to do, so I had to fix it! I made some annotations on the first heart I printed on the glass above -then I wiped it off by accident.

I got the right shape. Then I tested the print height, speed, and pressure so as to get better lining. It was much easier with a camera that caught the entire printing process and a microscope that allowed me to look at heart carefully.

Almost ready! I printed another heart with multi-layers. I put the glass into the over to cure the structure. For some other inks such as PDMS or gelMA, I would use UV curing.

Because of the hurricane, the fireworks celebration in Boston was moved to July 3rd, so I went to the bank to see the fireworks after the lab. Unfortunately, the storm hit us 5 minutes after the fireworks were over. I was soaked in water - so was my backpack. It took two hours and a half for me to get back to my apartment... So I caught a cold.

Peace.


P.S.2: During Week 3 and 4, besides finishing up my tasks, coding, printing filaments of different materials, preparing and aliquoting all kinds of solutions, observing cell behavior with various kinds of microscopes, I also shadowed two post-docs and two interns when I was free. Their work was really impressive. So were the dinosaurs, castles and a bird that were printed out by the expert engineers!