Day 7

Today I mainly worked with Sarah to continue growing organoids in the tissue culture lab to be able to insert them into mice to observe their growth and test different drugs on them to observe their responses. Sarah and I finished up plating the organoids that were left and after that was finished, we began to log data (a lot of what we did was digital data analysis) for the organoids before testing her drug—labeled 119 on the mice. I also got to go with Dr. Hsu to order thirty mice for this next wave of experiments with them. Today was mainly a research and data analysis day to finish up any pre-requisite steps to testing the mice. But for the past few days, we’ve mainly been working on the same thing— growing organoids and plating them to be ready for the mice. The reason why we have to do this over several days is because organoid growth doesn’t occur slowly, but it doesn’t occur rapidly either. If we put the organoids in an environment where they would rapidly grow, they would be diluted with fibroblasts (I talked about these in a previous post), and thus the sample wouldn’t be usable for the purpose of our research outside of the mouse. So it’s a little bit of a lengthy and slow process while there’s not much we can do other than pre-lab data analysis.

Day 7

Today was an excellent day at Huff Orthopedics. After meeting in the parking lot of the hospital, we made our way to the locker room as usual. In regards to the surgeries, Dr. Huff completed another rotator cuff repair, a knee scope and a bursa mass resection on the elbow.

In the first surgery, the rotator cuff repair, Dr. Huff used the shoulder scope device to move around inside of the shoulder joint. He then used a suture along with anchors to secure the tear in the cuff for recovery.
The second surgery consisted of a meniscus repair on the knee joint. In the surgery, Dr. Huff used the knee scope devices. These devices are very similar to the shoulder scoping equipment. It was amazing to me because this surgery only lasted ten minutes.

The final surgery consisted of Dr. Huff using a scalpel to cut a bursa mass from the elbow. This mass was most likely from a fall when the patient hit their elbow in the past. The surgery was fairly simple, Dr. Huff cut into the skin to reach the mass which he then cut out. However, he had to be especially careful for hitting a nerve which controls the hand.

To wrap up the day, Dr. Huff and I met with many patients at his office. Overall, I have had a fantastic week, and I look forward to my final day at the office!

Day 7 – Filing Day

Today was a slower day around the office. I worked some more on finding prospects and events to get into contact with, and made some phone calls to clients that won concert tickets. Around 11:30 I was tasked with going through and organizing the filing cabinets!!!! It was amazing to see all of the things that accumulated over the years, and I got to learn more about the accounts of sponsors and the importance of keeping almost all records on file (always keep W9 forms, I don’t know what they are used for, but Maria said they are really important so never throw yours away). On slower days it is harder to appreciate the work that I am doing, but it has truly given me a look into what a true office day is like and it has taught me that not every day is going to be super eventful but it is easy to appreciate a break from all the madness of concerts!

Please enjoy this picture of a painting I took at Meymandi Concert Hall yesterday.

Day 7: Filing and Documenting

Today was another day at the KDB office. Today I sat in on another meeting, this time with some Intel executives, in which we discussed various upgrades to the technology of Intel Corporation. The meeting was very interesting I then shadowed Ms. Moye as she filed some documents and sent them off to Richard Dyer, an attorney for KDB. I spoke with John Kavinsky about patents that are currently pending and he even let me submit a patent to the USPTO myself, after I proofed it of course. I then saw how applications and declarations are filed. Everything was generally pretty straightforward, just a lot of copy and pasting. But tomorrow I get to speak with more attorneys and learn how their lives work!

Day 7 – Parlodion Magic

Today was full of learning new techniques and exciting experiments. Dr. Bermek and I spent the first half of the morning discussing the experiments she still had to complete before her paper about the Herpes Simplex virus could be written. We decided to perform an experiment that could potentially lead to never seen before results. There is a shape of DNA that people have not been able to get, but with the protein UL8 we have been working with, Dr. Bermek wondered if this protein could help make the DNA shape because UL8 relaxes DNA and creates a nicked structure. Dr. Bermek wasn’t super optimistic we would get results but she wanted to try anyway. A large experiment like the one we were about to do calls for a lot of controls because, if it works, we want to make sure we know exactly what helped create the desired DNA shape. We set up 7 reactions that each included or didn’t include a specific ingredient. For example, one sample didn’t have UL8 but had UL30. The reason is because there has been speculation that UL30 might cause DNA to become nicked, so we wanted to have it as an option, but if it worked, we wanted to make sure that it was the UL30 protein that caused the DNA shape not the UL8.

Dr. Bermek had me make the master mix which is basically a large volume of ingredients that needed to be added to each reaction. It is used to save time and eliminate unwanted errors. I was happy she trusted me enough to make it. The timing of the preparation of the samples was overwhelming and difficult. We had four different timers going all for different things. We had to add specific ingredients to some of the seven samples but not others in addition to making sure the timing was perfect. Some samples had to be heated for five minutes others had to be heated for 30, and there were other parts of the experiment that required keeping a close eye on the time. Dr. Bermek and I had a good system going to help everything run smoothly. She would take the samples out the hot water bath and give them to me so I could pipette the ingredients in. She would then put the samples back in the hot water bath to continue heating them. While she was working with radioactive materials, I prepared two Agarose gels. I measured out the Agarose, added the TAE or TBE buffer, dissolved the Agarose for 2 minutes and 15 seconds, let the mixture cool, and then poured the gel liquid into the apparatus to let it settle. This time, Dr. Bermek loaded the samples into the gel because some of them were radioactive and required special precautions. I didn’t trust myself enough to load after we had spent about 3 hours preparing the samples.

While the gel was running we prepared grids for the EM that we would use tomorrow. When I first used grids last week, they were already prepared so I was glad I got to see the full process today. There was a special solution we poured over the mesh where our grids would be housed. Once the mesh was completely submerged in the liquid, we carefully placed the grids one by one on the mesh making sure they stuck. I got to help Dr. Bermek with this step, and I really liked the precise nature of it. Then came the coolest part: adding the Parlodion. Parlodion is used to coat the gel grids so they can be used in the EM. The coolest thing was when we added a few drops of it to the solution that the mesh was in, it created a rainbow colored layer over the grids. I was really fascinated.

By the time we had finished the UL8 experiment and set up the grids, they day was just about over. Even though we may not get the results that weren’t likely to happen, I’m excited to see what the gel image and EM show us tomorrow on my last day working with Dr. Bermek.

The gel that contains radioactive material getting run behind a protective shield.

Getting ready to add the Parlodion.

The cool rainbow effect of the Parlodion.

The Agarose gel I set up.

Our designed experiment plans for the day.

Day 7: Visit and Variations

It’s hard to believe that today was my second to last day at LCA! I have been having such an unforgettable and educational experience, having gained so much exposure to a potential area of college study and life-long profession. Shoutout to Ms. Eason for paying me a visit today; it was nice to see a familiar face in the office. I made some great progress on my greenhouse today. I completed the frame of my initial design, which made my next step of testing out glass paneling easier. By having a fundamental frame, I could copy and paste that same frame, only changing the glass for each variation. The traditional greenhouse has sectioned glass that usually follows a rectangular shape. I tested out several options regarding the sizes of the glass panels—big or small rectangles, horizontal, and vertical. There is no correct way to go about designing this greenhouse; it is important to test out the many different designs that can be created so the best one can be chosen. I am excited for my last day tomorrow but also a little nervous! The designers and I are having a group critique with the various projects we have all been working on. In the critique, I will be able to experience a true ‘design-thinking’ environment. Wish me luck!

Work Experience Day 7- Super Scanning and File Finding

Every day I have spent at the Polanco firm has been extremely interesting, I have loved getting to learn more about this type of career. and getting to work alongside amazing people. Today, I started my morning scanning lots of papers. It took me a long time to get used to it, the scanner was very finicky. Occasionally it would malfunction (or I would put the papers in crooked) which would results in a slightly embarrassing and very loud error alert. However, I soon got the hang of it. I also became adept at removing staples from papers so they could be put through the scanner. Even though this task sounds (and was at times) monotonous, it was cool to be involved in such an important process. A person’s immigration story is wrapped up in their file. Their file gives all the facts about their life, including their name, language, family, country of origin, and occupation. They show their story through a lawyers scraggly notes in messy handwriting, copies of visa applications, printed out copies of exchanged emails, and their family member’s written requests for help. So, I loved being part of this process even if it was just through making copies. Throughout the day, I also spent more time entering data online, updating closed case file boxes, and preforming other small errands. I am very proud of myself that I have learned which file boxes hold names starting with each letter of the alphabet and that I have become skilled at knowing what to look for when it comes to data entry. One of my most shining moments today came when I was handed a stack of about 10 receipts to file. Usually when filing, there are a lot of papers left over because it can be almost impossible these files. At times, it seems like we are looking for needles in a haystack. But today, I was extremely proud that I was able to file all these papers in record time! I was really glad I was able to help out so effectively without needing to stop others in their work to ask for aid.

What was probably the most exciting part of my day, however, came when Mrs. Eason came to visit the firm. While I was talking with her and the person who had been overseeing me throughout this week, the Polanco law employee mentioned how grateful she was that I was there to help out. She mentioned that she didn’t want me to leave and said I was welcome back anytime. I was so glad that I had been helpful at a firm that does so much good in the world and am excited to say that I will probably come back to work with them some this summer! Overall, this was another great day at the law firm and I cannot wait to work with them more.

Day 7 – Successful Troubleshooting

My visit to Chesterfield was an exciting one today. I worked with Natalia, a PHD student who explained to me various forms of 3D Printing and their pros and cons. See the photo below for a detailed chart of the main different types. For reference, I have worked with Material Extrusion (ultimaker), powder bed fusion (titanium printer), and vat photopolymerization (the carbon printer). Natalia explained how different printers have different tendencies to create structural defects and she is researching how to address and minimize these inconsistencies. For example, material extrusion printers result in printing by layer, so the x and y axis of the structure are relatively strong, but the z-axis may be weaker since the space between layers allows structural weakness. Natalia is testing what settings would be most optimal for printers to minimize this decrease in “weld strength.”

Natalia is currently working with three different materials: PCU (Polycarbonate urethane), the flexible but fairly strong polymer I worked with Friday, PEEK (Poly ether ether ketone) an crysalline polymer, and PEKK (poly ether ketone ketone), an amorphuous polymer. The latter two materials are unique in that they require much higher temperatures than the average 200*C to melt. Normal material extrusion printers can’t attain a temperature that high on their nozzles, so a special material extrusion printer with an enclosure and extra heated build plate is required to print PEEK and PEKK, going up to 450*C. Unfortunately, this printer had some teflon stuck in the spool feeder, so we had to put the whole tube in the oven at 190*C to melt it out. Luckily, we succeeded and reinstalled the tube into the printer after a an hour or two of heating!

After explaining the different material types, Natalia gave me a more detailed tour of the lab. One interesting contraption is the extruder that creates printable materials. Normally, a spool of material of a specified diameter (1.95 or 2.85 mm) is used for material extrusion printing (see the provided image for reference). However, manufactures may not produce materials that come in spools; rather, they come in pellet form. This extruder machine converts these pellets into a usable spool of printing material by melting the material and reforming it into a thread. It’s a shame, however, that the machine is currently broken since the extrusion tip is clogged. The lab members are working on fixing it, but they have no idea what is wrong after multiple attempts to follow standard protocol. In theory, the machine is highly useful for producing spools of important materials that you can’t buy of the shelf in spool form.

Throughout the lab, there are also other components that need fixing. We looked at the Lulzbot, a material extrusion printer with a twist; it has a flexistrude tip that allows for more accurate extrusion since the motor that moves the spool is closer to the tip. On this contraption, Natalia and I worked on fixing one of the resistors that heats up the nozzle, as well as the temperature indicator on the device. Meanwhile, we printed a cylinder of PCU on a second Lulzbot; this cylinder will later be cut with the laser cutter into a dog bone for mechanical testing, similar to what I did on Friday with Will. After a failed attempt and some more troubleshooting, the print was on its way to be done! One fun fact about material extrusion printing that we took into account while printing the cylinders is: a skirt (small ring around object) is required to help with inconsistencies in the extrusion and may also provide support for the structure.

Overall, today was a fun and educational day! Natalia was very friendly and explained the project well – I hope I can learn even more from her tomorrow. Hopefully the last day will be the best one yet!

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Day 7 – IVRT

Today, we shadowed a few scientists at in vitro release testing (IVRT). To start off, Craig, the head of the IVRT team, talked us through some of the experiments performed and methods used by IVRT, including different dosing techniques and strengths of drugs and how they impact the release rates. Then, Akeem demonstrated a test using vertical diffusion cells. The setup was similar to what we saw yesterday, simulated the properties of skin with a synthetic membrane rather than using actual skin, requiring a smaller time frame for each experiment. In order to determine the release medium that would result in an optimal dose-release ratio, Akeem tested two different solutions: a 50:50 mixture of water and methanol and a 70:30 ratio of water and ethanol. Rather than applying the cream with a pipet as the scientists in IVPT did, he added a constant volume of the drug to each cell by filling the hole in a washer-shaped wafer placed on top of the membrane.

After lunch, we talked with Meredith, who explained some of the functions of HPLC machines and what she looks for on the chromatographs. Then, Ms. Allen paid us a visit, and we discussed our experiences over the past two weeks. Finally, we met with Craig again; he discussed some more of the FDA regulations and guidelines that have to be followed when testing new proposed drugs, as well as the different types of tests performed on the drugs, some of which can take years to complete.

Day 7: Algae and Pig Farming

In the morning I worked with Elle and talked to her about different types of harmful algae. She was specifically working with Pfiesteria (algae that can be toxic and lead to fish kills) and let me see them under a microscope. She had samples from various locations in North Carolina as well as a few from New Zealand. After that, I helped her feed the algae and learned a lot about writing research reports and her old projects. Elle formerly worked on a project about using algae as a biofuel, which I found incredibly interesting. I also got a refresher on types of microscopes (since 9th grade I’ve forgotten a lot of biology haha). Some of the pictures below are ones she took while doing her various research projects.

Ms. Mackenzie and I went back to the Cary Wastewater treatment plant to pick up a sample of their inflow. It was a brief visit, but it was nice seeing one of the lab analysts for their lab (they weren’t there on the tour). Towards the end of my day, I watched Ms. Mackenzie do a test for fecal coliform (it involves pretty red and blue solutions).

I also had the privilege to speak with Dr. JoAnn Burkholder for about an hour. She gave me a brief presentation about water quality in North Carolina and how government policy is needed to genuinely improve it. North Carolina is one of the top pig farming states and is also competitive in the poultry industry; thus making animal waste a big issue. Due to a lack of regulations, this waste is often carelessly disposed of and negatively impacts communities and waterways surrounding the farms.

Image 1: Sample from water treatment plant

Image 2: Pfiesteria in bottles (pink coloring is because of their food)

Image 3: Fecal coliform test

Image 4: Pfiesteria piscicida zoospore (in the bottles)

Image 5: “Angry face anabaena” – Anabaena planctonica (picture from one of Elle’s previous projects)