Browsing by Subject "Optical imaging"

Now showing 1 - 4 of 4
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    Author Correction: Super-resolution microscopy compatible fluorescent probes reveal endogenous glucagon-like peptide-1 receptor distribution and dynamics
    (Nature Publishing Group, 2020-10-09) Ast, Julia; Arvaniti, Anastasia; Fine, Nicholas H. F.; Nasteska, Daniela; Ashford, Fiona B.; Stamataki, Zania; Koszegi, Zsombor; Bacon, Andrea; Jones, Ben J.; Lucey, Maria A.; Sasaki, Shugo; Brierley, Daniel I.; Hastoy, Benoit; Tomas, Alejandra; D’Agostino, Giuseppe; Reimann, Frank; Lynn, Francis C.; Reissaus, Christopher A.; Linnemann, Amelia K.; D’Este, Elisa; Calebiro, Davide; Trapp, Stefan; Johnsson, Kai; Podewin, Tom; Broichhagen, Johannes; Hodson, David J.; Pediatrics, School of Medicine
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    Editorial: Pancreas Imaging Across the Spectrum
    (Frontiers Media, 2021-01-03) Linnemann, Amelia K.; Poitout, Vincent; Rutter, Guy A.; Pediatrics, School of Medicine
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    Multiplexable fluorescence lifetime imaging (FLIM) probes for Abl and Src-family kinases
    (Royal Society of Chemistry, 2020) Jena, Sampreeti; Damayanti, Nur P.; Tan, Jackie; Pratt, Erica D.; Irudayaraj, Joseph M.K.; Parker, Laura L.; Neurological Surgery, School of Medicine
    Many commonly employed strategies to map kinase activities in live cells require expression of genetically encoded proteins (e.g. FRET sensors). In this work, we describe the development and preliminary application of a set of cell-penetrating, fluorophore labelled peptide substrates for fluorescence lifetime imaging (FLIM) of Abl and Src-family kinase activities. These probes do not rely on FRET pairs or genetically-encoded protein expression. We further demonstrate probe multiplexing and pixel-by-pixel quantification to estimate the relative proportion of modified probe, suggesting that this strategy will be useful for detailed mapping of single cell and subcellular dynamics of multiple kinases concurrently in live cells.
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    Super-resolution microscopy compatible fluorescent probes reveal endogenous glucagon-like peptide-1 receptor distribution and dynamics
    (Nature Research, 2020-01-24) Ast, Julia; Arvaniti, Anastasia; Fine, Nicholas H. F.; Nasteska, Daniela; Ashford, Fiona B.; Stamataki, Zania; Zania, Zsombor; Bacon, Andrea; Jones, Ben J.; Lucey, Maria A.; Sasaki, Shugo; Brierley, Daniel I.; Hastoy, Benoit; Tomas, Alejandra; D’Agostino, Giuseppe; Reimann, Frank; Lynn, Francis C.; Reissaus, Christopher A.; Linnemann, Amelia K.; D’Este, Elisa; Calebiro, Davide; Trapp, Stefan; Johnsson, Kai; Podewin, Tom; Broichhagen, Johannes; Hodson, David J.; Pediatrics, School of Medicine
    The glucagon-like peptide-1 receptor (GLP1R) is a class B G protein-coupled receptor (GPCR) involved in metabolism. Presently, its visualization is limited to genetic manipulation, antibody detection or the use of probes that stimulate receptor activation. Herein, we present LUXendin645, a far-red fluorescent GLP1R antagonistic peptide label. LUXendin645 produces intense and specific membrane labeling throughout live and fixed tissue. GLP1R signaling can additionally be evoked when the receptor is allosterically modulated in the presence of LUXendin645. Using LUXendin645 and LUXendin651, we describe islet, brain and hESC-derived β-like cell GLP1R expression patterns, reveal higher-order GLP1R organization including membrane nanodomains, and track single receptor subpopulations. We furthermore show that the LUXendin backbone can be optimized for intravital two-photon imaging by installing a red fluorophore. Thus, our super-resolution compatible labeling probes allow visualization of endogenous GLP1R, and provide insight into class B GPCR distribution and dynamics both in vitro and in vivo.