Lab Experience at the Department of Infectious Diseases and Tropical Medicine of San Paolo Hospital, Milan – Karol Jacek Kieszkowski

I got the opportunity to do my 1st lab rotation at the Laboratory of Professor Giulia Marchetti, Department of Infectious Diseases and Tropical Medicine of  San Paolo Hospital, University of Milan, Milan, Italy.

PI of the laboratory: Prof. Giulia Marchetti

Tutor: Dr. Roberta Rovito

The lab focuses on studying the immunopathogenesis of different viral infections, such as SARS-CoV-2 and HIV, by means of different techniques, such as flow cytometry. However, for my lab rotation we focused on the immunopathogenesis of long-COVID, i.e. the long-term persistence of debilitating symptoms even after the resolution of SARS-CoV-2 infection.

My experience & learnings:

During my lab rotation, I have learnt many useful research procedures spanning the entire process, i.e. from the biological specimen isolation to data analysis and results interpretation. In particular:

• Peripheral blood mononuclear cells isolation by means of density gradient centrifugation;
• Plasma and serum isolation;
• Cell thawing;
• Antibody titration;
• Basics on the data acquisition by CYTEK Aurora spectral flow cytometer;
• Basics on the data analysis of acquired flow cytometry data by the FlowJo software, including gating of cell populations and calculation of the stain index. It is needed to find the optimal antibody titre for flow cytometry experiments;
• Ability to utilize relevant sources for ongoing research projects undertaken by the team.
• Wide knowledge about the pathogenesis of the long COVID: one of the main subjects studied by the Infectious Diseases team.

The internship was challenging, considering my lack of previous experience in the laboratory. However, I was provided with relevant resources, such as the papers published by the team that I followed, which greatly helped me understand the main topic of their current studies – the Long COVID. Moreover, I performed all of the experimental procedures, especially in the first days of rotations, under the strict guidance of my tutor. She provided me with protocols for each undertaken procedure in advance, which really helped me prepare for each day of my rotation. In addition, the whole team was always eager to explain me any concept that I found difficult.

The rotation helped me develop scientific reasoning. In addition to the tasks in the lab, I was involved in a second research project. In particular, my role was to search the medical literature for information that would help designing experiments to study specific cellular pathways possibly altered in long COVID based on previous transcriptomic results of the lab. This process was a valuable opportunity for me to learn how to find relevant sources, which keywords to use, how to critically read a scientific paper, and which sections of the article to focus on in the context of the project.

I am now equipped with valuable skills and knowledge that will surely help me in my future academic activities especially in other lab rotations and  my work on the experimental thesis. Moreover, I believe that the ability to find up-to-date academic resources be useful for me not only in research, but also in my every day medical studies.