Light scattering was recently demonstrated to serve as an intrinsic indicator for pancreatic islet cell mass and secretion. The insulin secretory granule (ISG), in particular, was proposed to be a reasonable candidate as the main intracellular source of scattered light due to the densely-packed insulin semi-crystal in the granule lumen. This scenario, if confirmed, would in principle open new perspectives for label-free single-granule imaging, tracking, and analysis. Contrary to such expectations, here we demonstrate that ISGs are not a primary source of scattering in primary human β-cells, as well as in immortalized β-like cells, quantitatively not superior to other intracellular organelles/structures, such as lysosomes and internal membranes. This result is achieved through multi-channel imaging of scattered light along with fluorescence arising from selectively-labelled ISGs. Co-localization and spatiotemporal cross-correlation analysis is performed on these signals, and compared among different cell lines. Obtained results suggest a careful re-thinking of the possibility to exploit intrinsic optical properties originating from ISGs for single-granule imaging purposes.
Probing the light scattering properties of insulin secretory granules in single live cells
Bugliani, Marco;Marchetti, Piero;
2018-01-01
Abstract
Light scattering was recently demonstrated to serve as an intrinsic indicator for pancreatic islet cell mass and secretion. The insulin secretory granule (ISG), in particular, was proposed to be a reasonable candidate as the main intracellular source of scattered light due to the densely-packed insulin semi-crystal in the granule lumen. This scenario, if confirmed, would in principle open new perspectives for label-free single-granule imaging, tracking, and analysis. Contrary to such expectations, here we demonstrate that ISGs are not a primary source of scattering in primary human β-cells, as well as in immortalized β-like cells, quantitatively not superior to other intracellular organelles/structures, such as lysosomes and internal membranes. This result is achieved through multi-channel imaging of scattered light along with fluorescence arising from selectively-labelled ISGs. Co-localization and spatiotemporal cross-correlation analysis is performed on these signals, and compared among different cell lines. Obtained results suggest a careful re-thinking of the possibility to exploit intrinsic optical properties originating from ISGs for single-granule imaging purposes.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.