Photostable and Orthogonal Solvatochromic Fluorophores
for Simultaneous In Situ Quantification of Multiple
Cellular Signaling Molecules
Posted on 2020-06-25 - 14:35
Ratiometric
fluorescence sensors are powerful tools for direct
quantification of diverse biological analytes. To overcome a shortage
of solvatochromic fluorophores crucial for in situ ratiometric imaging of biological targets, we prepared and characterized
a small library of modular fluorophores with diverse spectral properties.
Among them, WCB and WCR showed excellent spectral properties, including
high photostability, brightness, and solvatochromism, and are ideally
suited for dual ratiometric imaging due to their spectral orthogonality.
By conjugating WCB and WCR with protein-based lipid sensors, we were
able to achieve robust simultaneous in situ quantitative
imaging of two metabolically linked signaling lipids, phosphatidylinositol-4,5-bisphosphate
and phosphatidylinositol-3,4,5-trisphosphate in live cells. This study
shows that any combination of signaling molecules can be simultaneously
quantified in a spatiotemporally resolved manner by ratiometric imaging
with finely tuned solvatochromic fluorophores.
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Sharma, Ashutosh; Sun, Jiachen; Singaram, Indira; Ralko, Arthur; Lee, Daesung; Cho, Wonhwa (2020). Photostable and Orthogonal Solvatochromic Fluorophores
for Simultaneous In Situ Quantification of Multiple
Cellular Signaling Molecules. ACS Publications. Collection. https://doi.org/10.1021/acschembio.0c00241