Year: 2026
Type: article
Abstract: ABSTRACT The non‐covalent interactions that underpin major cellular functions depend on molecular motion within 3D environments. Large depth‐of‐field single‐molecule localization microscopy (3D‐SMLM) methods facilitate these measurements, but their increased optical complexity and bespoke post‐processing pipelines often sacrifice important cellular context. Here, we combine single‐molecule light‐field microscopy (SMLFM) with widefield Fourier light‐field microscopy for correlative volumetric organelle imaging. The instantaneous acquisition of subcellular volumes improves the sensitivity of molecular organization, chemical environment, and diffusion measurements through the use of volumetric sub‐cellular segmentation. We first demonstrate our approach by measuring the molecular organization of a nuclear‐localized HaloTag protein relative to cell nuclei. Next, we characterize the molecular diffusion of the soluble protein, calreticulin, in the context of ‐antitrypsin deficiency, which revealed an increase in heterogeneous motion within endoplasmic reticulum inclusions.
Source: Small
Institutions: University of Cambridge, MRC Laboratory of Molecular Biology
Language: English
Subfield: Biophysics
Domain: Life Sciences
Open Access status: hybrid
Estimated APC paid: $5,790
Awards: Linking alpha1-antitrypsin phase transition with cellular health, G1810828351, G2406928993, BBSRC IAA University of Cambridge, How does the Golgi sort proteins? Mapping the Biophysical Landscape of the Secretory Pathway, Effects of alpha1-antitrypsin polymerisation on organelle structure and fluidity in hepatocytes, G9000229773
OpenAlex ID: W7124265114