Stimulated Raman Scattering (SRS) microscopy is a powerful nonlinear optical microscopy technique that images biological structures by exploiting the characteristic, vibrational contrast of the sample molecules. SRS provides a rich, chemically specific and biophysical image contrast that is in many ways complementary to the molecular contrast of fluorescence microscopy. Here, we present a range of applications of SRS, including label-free morphochemical imaging in model organisms, the characterization of organoids and spheroids, and investigations of brain tissues for neurodegenerative disease research. We show specifically that SRS can provide novel insights into the biophysical properties and biochemical composition of Amyloid-β plaques in a mouse model of Alzheimer’s disease. Our results highlight the potential of SRS to contribute to a deeper understanding of cell and tissue biology, and to serve as a powerful tool for preclinical and translational research.
The precise sub-cellular spatial localization of multi-protein complexes is increasingly recognized as a key mechanism governing the organization of mammalian cells. Consequently, there is a need for novel microscopy techniques capable of investigating such sub-cellular architectures in comprehensive detail. Here, we applied a novel multiplexed STORM super-resolution microscopy technique, in combination with high-throughput immunofluorescence microscopy and live-cell imaging, to investigate the roles of the scaffold protein
IQGAP1 in epithelial cells. IQGAP1 is known to orchestrate a wide range of biological processes, including intracellular signaling, cytoskeletal regulation, cell-cell adhesion, and protein trafficking, by forming distinct complexes with a number of known interaction partners, and recruiting these complexes to specific subcellular locations. Our results demonstrate that, in addition to supporting epithelial adherens junctions by associating with specialized cortical actin structures, IQGAP1 plays a second role in which it controls the confinement of a unique, previously undocumented class of membranous compartments to the basal actin cortex. These largely immotile yet highly dynamic structures appear transiently as cells merge into clusters and establish of apical-basolateral (epithelial) polarity, and are identified as an intermediate compartment in the endocytic recycling pathways for cell junction complexes and cell surface receptors. Although these two functions of IQGAP1 occur in parallel and largely independently of each other, they both support the maturation and maintenance of polarized epithelial cell architectures.
Conference Committee Involvement (1)
Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XIX
6 March 2021 | Online Only, California, United States