The pitch lab prep

Tell me about yourself.

I’m a rising junior at Tufts studying Biopsychology. My research experience has centered on genetics, neurobiology, and fluorescence imaging. I first completed a year-long Drosophila genetics project in which I generated and characterized a novel enhancer-trap line. This summer, I’ve been contributing to an ongoing project in the Dulla Lab characterizing atypical astrocytes in APP mouse models of Alzheimer’s disease. I’m now looking for a longer-term position where I can build on my imaging and analysis experience while taking greater ownership of a defined project.

How would you describe your research interests, and how did you develop them?

I’m broadly interested in how cellular and molecular changes in the nervous system lead to disease-related dysfunction. I first became interested in this through my Drosophila project, where I saw how a genetic change could be connected to a specific expression pattern in the nervous system. In the Dulla Lab, I became more interested in neurodegeneration and in how cellular phenotypes can be identified through molecular markers and imaging. I’m especially drawn to projects that connect cell biology to disease mechanisms and potential therapeutic questions. I’m still refining my specific interests, but I know I want to understand how a biological question progresses from an initial observation through experiments, analysis, and publication.

What have you been working on in the Dulla Lab?

I’ve been contributing to my mentor’s larger project characterizing atypical astrocytes in APP mouse models of Alzheimer’s disease. My role has mainly involved mouse and tissue preparation, immunohistochemistry, fluorescence imaging, antibody validation, and a defined ImageJ analysis. My clearest analytical contribution was comparing background-corrected Iba1 fluorescence across atypical astrocytes, typical astrocytes, and microglia as part of evaluating the cellular identity of the atypical astrocytes. I’m also beginning work examining PV, GAT1, and GAT3 in relation to ATAs in the thalamus.

What was your specific contribution to the Iba1 analysis?

I identified atypical astrocytes using regions of GLAST signal dropout, selected the relevant cellular regions, measured local background fluorescence, and performed background-corrected comparisons of Iba1 signal among ATAs, typical astrocytes, and microglia. The analysis was part of a larger effort to determine whether ATAs are molecularly distinguishable from other cell populations. I contributed to the quantification, but I did not design the overall study.

What other techniques have you used in the Dulla Lab?

I’ve maintained mouse inventory, handled mice, performed transcardial perfusions, extracted brains, cryosectioned tissue, carried out immunohistochemistry, participated in RNAscope workflows, mounted tissue, acquired multichannel fluorescence images, and used ImageJ for quantitative analysis. I’ve also tested antibody-staining conditions by comparing different concentrations and protocols with and without antigen retrieval.

What do you mean when you say you validated antibodies?

I followed immunohistochemistry protocols using different antibody concentrations and, in some cases, compared staining with and without antigen retrieval. The goal was to identify conditions that produced strong, interpretable target signal while minimizing background and nonspecific staining. I carried out the experimental comparisons using protocols planned with my mentor rather than independently designing the entire validation strategy.

Can you describe your ImageJ experience?

I’ve used ImageJ for ROI-based fluorescence analysis in multichannel images. In the Iba1 analysis, I identified ATAs using GLAST dropout regions, measured background signal, and compared background-corrected fluorescence across cellular groups. I understand that mean intensity measures average signal within an ROI, while integrated density is affected by both intensity and ROI size. My current experience has mostly involved following an established analysis workflow, and I’m interested in becoming more independent in designing and validating complete image-analysis pipelines.

What is your experience with confocal microscopy?

What is your experience with fluorescence and confocal microscopy? I have the most independent experience with the Leica THUNDER fluorescence-imaging system. I’ve adjusted exposure for individual channels, selected objectives and channel settings to reduce bleed-through, established imaging boundaries, created focus maps across tissue sections, and assigned display colors to different markers. I understand the importance of avoiding saturation and maintaining consistent acquisition settings when fluorescence will be compared quantitatively. I have also used the Leica FALCON confocal microscope, but mainly with acquisition settings established by my mentor. I understand its basic principles, including optical sectioning, rejection of out-of-focus light, multichannel imaging, and z-stacks, but I have less experience independently optimizing that system.

Tell me about your Drosophila research project.

I completed a year-long enhancer-trap project in Drosophila. I generated and stabilized a novel line through genetic crosses and balancer chromosomes, maintained the stocks, and sorted offspring to preserve the desired insertion. I then used inverse PCR and sequencing to identify the genomic DNA adjacent to the enhancer-trap insertion, which allowed us to map it upstream of the serrano gene. I also dissected larval central nervous systems and used confocal microscopy to characterize reporter expression, including expression in the mushroom bodies and tracheal branches. The line was stabilized and archived for future work.

What did the Drosophila project teach you?

It taught me how different techniques can answer different parts of the same biological question. The genetic crosses established and stabilized the model, inverse PCR and sequencing identified where the insertion occurred, and confocal imaging showed where the associated regulatory element was active. It also taught me patience, because maintaining the line and obtaining the correct crosses required work across many generations.

Do you have cell-culture experience?

I have not independently maintained mammalian cell cultures. My current work has primarily involved fixed mouse brain tissue rather than living cultured neurons. However, I’m accustomed to detailed multistep protocols, delicate biological samples, time-sensitive experiments, microscopy, and careful documentation. I understand that cell culture requires aseptic technique, consistent maintenance, monitoring cell health, and coordinating experiments with culture age, and I’m prepared to learn those practices carefully until I can work independently.

Why are you interested in this particular project?

I’m interested because it asks whether a visible cellular feature—endo-lysosomal size—corresponds to meaningful functional differences. That connects well with my experience using fluorescence measurements to compare cellular populations, but it would also introduce me to live-neuron imaging and cell culture. I’m especially excited that the project has a defined remaining scope and could allow me to take responsibility for completing experiments, analyzing the data, and participating in the manuscript process.

Why are you looking for another lab if you are already in the Dulla Lab?

My experience in the Dulla Lab has been valuable for building technical skills in tissue processing, immunohistochemistry, imaging, and quantification. However, my role has mainly involved carrying out assigned parts of my mentor’s larger project rather than understanding or directing a defined project myself. I’m looking for a long-term opportunity where I can become more involved in the scientific reasoning, take responsibility for a specific project, and follow it through analysis and publication.

What are your main strengths as a researcher?

I’m careful with multistep protocols, I’m comfortable working with fluorescence images and quantitative measurements, and I’ve demonstrated that I can remain committed to a project over a long period. My Drosophila project also showed that I can connect genetic, molecular, and imaging results rather than treating each technique as an isolated task. I’m also honest about what I know and what I still need to learn, which I think is important for becoming reliable in a new experimental system.

What is one weakness or area you want to develop?

I want to become more independent in understanding experimental design and building analysis workflows rather than only following established instructions. I also want to develop cell-culture and live-cell imaging experience. I see those as important next steps because I want to progress from being technically reliable to being able to understand, troubleshoot, and contribute intellectually to an entire project.

How have you grown this summer?

One of the biggest areas of growth for me this summer has been learning how to make myself reliable in a laboratory setting. At the beginning, I sometimes relied too heavily on remembering verbal instructions, which meant I occasionally needed steps repeated. I realized quickly that this would limit my ability to become independent, so I developed a much more systematic approach.

Now, I take detailed notes not only on the steps of a protocol, but also on timing, reagent locations, common mistakes, and the reasoning behind important steps. After learning a procedure, I reorganize my notes into a format I can follow independently and update them whenever I receive feedback. That system has made me much more confident in my ability to reproduce procedures consistently without needing repeated instruction.