Huygens deconvolution has been featured in a recent publication in Scientific Reports.
Researchers from the University of Turku in Finland showcased the adoption of the specific hybridisation internalisation probe (SHIP) assay in studies on the trafficking of receptor/ligand complexes. In their research, Hernández‑Pérez and Mattila used B lymphocytes and B cell receptor‑mediated antigen internalisation as a model system.
Images acquired with a spinning-disk confocal microscope were deconvolved using Huygens Essential’s MLE algorithm prior to co-localisation analysis. The results were also contrasted against those of Stimulated emission depletion (STED) microscopy, Super-resolution radial fluctuations (SRRF), and Airyscan. The resolution of deconvolved spinning-disk confocal images was comparable to that of STED and SRRF-processed images, and superior to Airyscan.
Image from Figure 6D (doi: https://doi.org/10.1038/s41598-021-04544-6)
Citation
Hernández-Pérez, S., Mattila, P.K. A specific hybridisation internalisation probe (SHIP) enables precise live-cell and super-resolution imaging of internalized cargo. Sci Rep 12, 620 (2022). https://doi.org/10.1038/s41598-021-04544-6
Abstract
Facilitated by the advancements in microscopy, our understanding of the complexity of intracellular vesicle traffic has dramatically increased in recent years. However, distinguishing between plasma membrane-bound or internalised ligands remains a major challenge for the studies of cargo sorting to endosomal compartments, especially in small and round cells such as lymphocytes. The specific hybridization internalisation probe (SHIP) assay, developed for flow cytometry studies, employs a ssDNA fluorescence internalisation probe and a complementary ssDNA quenching probe to unambiguously detect the internalized receptors/cargo. Here, we adopted the SHIP assay to study the trafficking of receptor/ligand complexes using B lymphocytes and B cell receptor-mediated antigen internalization as a model system. Our study demonstrates the potential of the SHIP assay for improving the imaging of internalized receptor/ligand complexes and establishes the compatibility of this assay with multiple imaging modalities, including live-cell imaging and super-resolution microscopy.
