The last time I was at the lab, I was there for a short visit to discuss my senior project in November. Previously, though, I spent the last two summers working there, and so it was wonderful to return to Dr. Christiano’s lab team this past Monday.
Once again joining Dr. Perez-Lorenzo, I was brought up to speed with all the recent developments at the lab. We discussed some of the new results and findings, as well as some of the ongoing investigations in the microbiome, alopecia, and melanoma which I was part of. The remainder of Monday was spent planning our experiment in greater detail, reaching out to colleagues to locate the needed tissue samples for optimization, as well as finishing to split some remaining cell cultures for the lab.
Tuesday morning we attended a HIMC(human immune monitoring center)seminar in Columbia on Vectra Polaris and Codex — “next-gen high-parametric tissue imaging for digital pathology and tumor-immune microenvironment discovery.” This highlighted some of the immense progress made in the field in terms of imaging techniques and technology while advertising some of the new opportunities that were made available for us this year in the Immune Core at CUMC.
After having to take care of an emergency broken freezer situation, so specimens will not ruin, we found the needed tissues. Returning to the lab, I began reading several research articles to do some literature review and choose the tight junctions I will focus on in the gut. The entirety of the day Wednesday was spent on beginning to work on the optimization of the staining for our initial gut tissue samples.
On Thursday we had our lab meeting. These are weekly meetings during which one researcher presents their progress, and others ask critical questions and give suggestions. Following the meeting, I resumed my staining. After deparaffinizing the slides, I used a blocking solution to mask any non-specific bindings. With some more washes, I also added the primary antibodies to the slides and left them to sit overnight in our walk-in fridge room.
On Friday, after making some antibiotic water for the mice, which contained a lot of sucrose so that they would drink it without realizing the bitter antibiotic contents, I went on to continue the staining. Adding secondary antibodies and going through some staining procedures, the slides were mounted and set to dry. After looking at the slides under the microscope, it proved difficult to distinguish between cells. This could be due to the small concentration of secondary antibody that was used, or more likely because I had previously been using dab-peroxidase brown and am not used to looking at cells after they are treated with nova red. Either way, I will need to add some more control slides to the protocol next week and will re-examine the slides after they sit for the weekend.