Abstract
Biochemical changes in specific organelles underpin cellular function, and studying these changes is crucial to understand health and disease. Fluorescent probes have become important biosensing and imaging tools as they can be targeted to specific organelles and can detect changes in their chemical environment. However, the sensing capacity of fluorescent probes is highly specific and is often limited to a single analyte of interest. A novel approach to imaging organelles is to combine fluorescent sensors with vibrational spectroscopic imaging techniques; the latter provides a comprehensive map of the relative biochemical distributions throughout the cell to gain a more complete picture of the biochemistry of organelles. We have developed NpCN1, a bimodal fluorescence-Raman probe targeted to the lipid droplets, incorporating a nitrile as a Raman tag. NpCN1 was successfully used to image lipid droplets in 3T3-L1 cells in both fluorescence and Raman modalities, reporting on the chemical composition and distribution of the lipid droplets in the cells.
Highlights
Understanding the biochemical composition of cells and organelles is essential for understanding both physiological and pathological processes
Whilst the core structure is modified for different applications, naphthalimides have seen little use in fluorescence multimodal imaging probes and are yet to be functionalised for vibrational spectroscopy
Bioorthogonal Raman tags in the biological-silent region from 1800 to 2800 cm−1 have enabled the identification of organelles, though many approaches rely on resonance Raman (RR) [49], stimulated Raman scattering (SRS) [24], or surface-enhanced Raman spectroscopy (SERS) [50] to enhance the signal
Summary
Understanding the biochemical composition of cells and organelles is essential for understanding both physiological and pathological processes. The comprehension of the chemistry of cells is greatly aided by tools to both identify and monitor the chemistry of organelles in cellulo. Small molecule fluorescent probes have emerged as important tools for investigating cells. Fluorescent probes can respond to specific biochemical species or stain specific sites on the cell, while being imaged at a high spatial and temporal resolution [1]. Most fluorescent sensors can only target one or two analytes of interest and crosstalk can complicate the use of multiple fluorophores to investigate different species. Fluorescent probes are less useful for investigating the general chemical composition of the cell and must be combined with other techniques to gain a more complete picture of the biochemical environment
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