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Chemiluminescence Microscopy Single Cell Imaging Using Esterase Triggered 1; 2-Dioxetanes
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DOI：10.1039/D5AN00696A.png)
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Abstract
En 中文
Chemiluminescence microscopy using emerging 1; 2-dioxetane chemiluminescent probes is an attractive prospect for low background biological molecular imaging. Elimination of an external light source reduces autofluorescence; light scattering; and complications from fluorophore photo-bleaching; providing enticing advantages compared to fluorescence imaging. While bioluminescence microscopy with genetically modified luciferase expressing organisms has been successful; examples of chemiluminescence microscopy using 1; 2-dioxetanes are limited due to challenges with cell uptake; signal brightness; and microscopy apparatus that are not optimized for these molecules. To address this gap; we herein describe a home-built chemiluminescence microscope capable of imaging nanomolar concentrations of luminescent 1; 2-dioxetanes. Using acetoxymethyl ester functionalized 1; 2-dioxetanes; 2 and 3; which display uptake and retention into unmodified A549 cells for imaging intracellular esterase activity; we characterize the luminescence imaging of A549 human lung epithelial cells via chemiluminescence microscopy. This study provides a complementary chemiluminescent 1; 2-dioxetane and microscopy system; thereby introducing a generalizable framework for imaging a wide variety of clinically relevant samples.
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