OregonFluor enables quantitative intracellular paired agent imaging to assess drug target availability in live cells and tissues.
Lei G WangAntonio R MontañoJason R CombsNathan P McMahonAllison SolankiMichelle M GomesKai TaoWilliam H BissonDani A SzafranKimberley S SamkoeKenneth M TichauerSummer L GibbsPublished in: Nature chemistry (2023)
Non-destructive fluorophore diffusion across cell membranes to provide an unbiased fluorescence intensity readout is critical for quantitative imaging applications in live cells and tissues. Commercially available small-molecule fluorophores have been engineered for biological compatibility, imparting high water solubility by modifying rhodamine and cyanine dye scaffolds with multiple sulfonate groups. The resulting net negative charge, however, often renders these fluorophores cell-membrane-impermeant. Here we report the design and development of our biologically compatible, water-soluble and cell-membrane-permeable fluorophores, termed OregonFluor (ORFluor). By adapting previously established ratiometric imaging methodology using bio-affinity agents, it is now possible to use small-molecule ORFluor-labelled therapeutic inhibitors to quantitatively visualize their intracellular distribution and protein target-specific binding, providing a chemical toolkit for quantifying drug target availability in live cells and tissues.
Keyphrases
- small molecule
- induced apoptosis
- high resolution
- cell cycle arrest
- fluorescent probe
- water soluble
- protein protein
- signaling pathway
- endoplasmic reticulum stress
- emergency department
- mesenchymal stem cells
- reactive oxygen species
- nitric oxide
- cell death
- mass spectrometry
- quantum dots
- sensitive detection
- bone marrow
- photodynamic therapy
- single cell
- living cells
- highly efficient
- fluorescence imaging
- dna binding