Week 1 Blog: Setting the Reagents and Experiments

Hello Everyone! Welcome to the first official week of my Senior Project. While the end goal is to help transform drug delivery, the reality of Week 1 is a bit more grounded, centered on preparation, and focused on reading a lot of research papers. In research, you can’t just jump into experimentation. I have to ensure my materials are pure, ready to use, and stable. I have to ensure my calculations are precise and accurate, and in this specific case, I have to ensure my environment is sterile. Think of this week like the French culinary term “mise en place,” which means having all your ingredients measured, cut, and ready before you start cooking. 

What I did this week:

• Material Procurement: I successfully bought and prepared the core components for my experiments: 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) lipids and Rhodamine B, the fluorescent dye that will allow me to visualize the vesicles. I also prepared the aqueous ionic buffer solutions I need. I prepared 0.1 M solutions of Sodium Chloride (NaCl), Calcium Chloride (CaCl₂), Potassium Chloride (KCl), and dPBS. I also tested for contamination. 
• Lipid Stock Solution: After I prepared the primary lipid stock solution. This involved dissolving the DOPC and trace amounts of dye into a 2:1 Chloroform/Methanol mixture to create a 10 mg/mL concentration.
• Sterilization: Purity is essential in biology and chemistry. I spent a significant portion of the week autoclaving pipette tips and glass slides to ensure absolutely no contaminants interfere with the delicate self-assembly process.
• Safety Review: Since I am handling chloroform and other volatile organic solvents, my advisor made me review the Safety Data Sheets (SDS) and established the proper fume hood protocols and other safety procedures to remain compliant with lab safety standards.

I have also started developing the Standard Operating Procedure (SOP) for the fluorescence microscope and Dynamic Light Scattering. While I haven’t finalized the protocol yet, I am currently testing different exposure times and gain settings to determine the optimal configuration. Locking these settings in now is important so there are no potential confounding variables or changes in arbitrary units. It basically ensures that any future differences I observe in vesicle yield or uniformity are due to the solvents, not inconsistent imaging or testing. Next week, I will continue this Protocol Optimization and then run the first trial runs. Thanks for reading!

– Samahith