Hello! My name is Katie Pumayalli, and I’ve just wrapped up the third week of my six-week internship at the Ye Lab at the Scripps Institution in San Diego, California. Can’t believe I’m already halfway through! I’m working under the guidance of the amazing Cailynn Wang and Zhengyuan (Ben) Pang. Since starting on June 17th, I’ve completely fallen in love with the lab environment and the people. My mentors’ research focuses on utilizing a technique called clearing-assisted tissue click chemistry (CATCH) to track hallucinogenic drugs within brain tissue. CATCH works by making brain tissue slices transparent. This is achieved by removing lipids (fats) from the cells, while still preserving the overall cellular structure. This innovative technique allows for precise visualization of where the drugs bind within the brain. This week, my primary focus was finalizing my Click reaction and gaining more practice with antibody staining.
Washed brain tissue
The week kicked off with our usual lab meeting on Monday. Afterward, I focused on completing the washing process for the tissue samples I collected from the sacrificed mice last week. Once clean, I stained the tissue with a specific chemical that binds to the drug. This staining allows us to visualize the drug distribution within the tissue, highlighting areas of high concentration and any discernible patterns. Finally, I utilized the fume hood to prepare a chemical solution for the tissue incubation.
Different wells of liquids for different staining processes
Tuesday brought a fascinating seminar on microRNA in the lens of neurons, which Siena, a fellow intern, and I attended. In the afternoon, I tackled the final steps of the Click reaction on the tissue I harvested. After mounting the tissue, I had to wait with anticipation until Thursday to see if my efforts were successful.
Shaker in a room that is constantly 4C used for tissues.
Wednesday offered a welcome break, allowing me to delve deeper into the lab’s research on various drugs. This enhanced knowledge will prove valuable when I analyze drug-binding patterns later. Additionally, I prepared a staining solution by mixing the staining component with a buffer. This solution will be used to visualize synapses, the communication junctions between neurons, in specific tissue samples. We’ll then be able to determine if the drug interacts with these critical structures.
Imaging of the drug tissue. (Notice all of the light in the top right corner)
Imaging of the vehicle tissue. (Notice the dim top right corner)
Excitement filled the air as Thursday arrived, the day I’d learn the outcome of my week’s work. Thanks to Cailynn’s invaluable guidance, my Click reaction was a success! I eagerly examined the brain tissue images, clearly differentiating between the vehicle and drug-treated mice. To cap off the day, I completed the secondary staining on the tissue samples prepared on Wednesday.
Staining of a different part of the neuron.
Friday kicked off with another round of Click reactions, this time targeting a different brain region. I also replenished the detergent buffer for the brain tissue I harvested two weeks ago. This was fascinating! Witnessing the tissue’s transformation over time, gradually becoming clearer, was truly rewarding. To conclude the week, I reviewed the images of the tissue I’d been processing since Wednesday.
Last weekend, Siena and I embarked on a fun-filled adventure. We explored the USS Midway Museum, gaining valuable insights into how the military safeguards our nation. We then immersed ourselves in the vibrant culture of San Diego with a visit to Seaport Village.
Thank you for reading in and can’t wait for next week’s fun adventures!
Katie Pumayalli
Congrats on your fun learning!