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Ifrah, a rising junior studying biochemistry and biophysics & black studies at Amherst College, investigated immunofluorescent antibody validation in adipocytes for LipocyteProfilerV2.

Understanding how cells respond to metabolic disease is essential for developing better treatments for conditions like diabetes and cardiovascular disease. Adipocytes, or fat cells, play a central role in these diseases, yet the full range of their structural and functional changes remains poorly understood.BSRP allowed me to challenge myself in ways that pushed me personally and academically, while being surrounded by an incredibly supportive community. It was a space where I could grow as a scientist and as an individual, with mentors and a cohort who encouraged resilience and confidence. LipocyteProfiler is a high-throughput, image-based phenotypic profiling platform that links genetic and chemical perturbations to adipocyte cell states using fluorescently labeled organelle markers. The original LipocyteProfiler includes validated antibodies or dyes for mitochondria (MitoTrackerRed, deep red), nucleus (Hoescht, blue), cytoskeleton/membrane (Phalloidin/WGA, yellow), and lipid droplets (BODIPY, green). 

However, it omits several organelles that may carry important phenotypic signals. This project aims to develop LipocyteProfilerV2 by expanding organelle coverage to enable more detailed subcellular profiling. We began with immunofluorescence protocols to evaluate antibodies targeting previously unprofiled structures. We successfully validated the GM130 antibody, which robustly labels the Golgi body, confirming its specificity and appropriate subcellular localization in differentiated adipocytes. The next step involves testing more antibodies for other organelles such as the endoplasmic reticulum, lysosome, and peroxisomes. By incorporating new organelle markers such as the Golgi body into LipocyteProfiler, we increase the potential to detect subtle or previously unresolvable cellular changes in response to perturbations. Ultimately, this work enhances the biological resolution of LipocyteProfiler, enabling researchers to better map the molecular and morphological changes that underlie metabolic disease. This expanded profiling capacity may facilitate new discoveries in adipocyte biology and support the identification of novel therapeutic targets.

Project: LipocyteProfilerV2: a novel metabolic antibody panel for adipocyte profiling

Mentor: Elizabeth Burton, Metabolism Program