![]() Integrating high-throughput screening with sophisticated high-resolution, high-sensitivity microscopy analysis is challenging because it usually requires specialized instrumentation for sample manipulation 3, 4. ![]() However, many cellular/subcellular phenotypes are only visible under high-resolution microscopes, and compared to high-copy transgenes, endogenous levels of gene expression are often low and require high-sensitivity detection. In particular, recently advanced genome editing approaches make it easy to tag endogenous proteins for visualizing their spatiotemporal distribution in living multicellular organisms. On the other hand, these multicellular organisms can also be advantageous to elucidate the molecular basis of cellular/subcellular structures and their functions in the context of physiological cell–cell interactions. High-throughput genetic screens using stereoscopes have identified various genes that regulate animal development, physiology, and behavior in multicellular organisms, such as Drosophila, C. The ability to study whole organisms makes it possible to study complex in vivo processes under physiological conditions that cannot be replicated in vitro or in cell culture systems. Together these studies demonstrate a photo-highlighting microscopic strategy for genome-scale screens, leading to the discovery of new roles for BMP signaling in linking mitochondrial homeostasis and lipid metabolism. Furthermore, this increase is associated with the induction of mitochondrial β-oxidation and mitochondrial fusion. Using SRS-based isotope tracing assay to quantitatively track lipid synthesis and mobilization, we discover that the BMP signaling mutants have increased rates of lipid mobilization. Four of these mutants target the components of the Bone Morphogenetic Protein (BMP) signaling pathway, revealing that BMP signaling inactivation causes exhaustion of lipid reserves in somatic tissues. We apply this method to stimulated Raman scattering (SRS) microscopy and systematically identify 57 Caenorhabditis elegans mutants with altered lipid distribution. ![]() Here we present a genetic screening method using photo-highlighting for candidate selection on microscopes. ![]() High-throughput screens at microscopic resolution can uncover molecular mechanisms of cellular dynamics, but remain technically challenging in live multicellular organisms. ![]()
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