Abstract
Fluorescent optical probes have rapidly transformed our understanding of complex biological systems by providing specific information on biological targets in the natural living state. However, their utility is often limited by insufficient brightness, photostability, and multiplexing capacity. Here, we report a conceptually new optical probe, termed ‘reflectophore’, which is based on the spectral interference from a dielectric microsphere. Reflectophores are orders-of-magnitudes brighter than conventional fluorophores and are free from photobleaching, enabling practically unlimited readout at high fidelity. They also offer high-degree multiplexing, encoded in their optical size, which can be readily decoded through interferometric detection with nanoscale accuracy, even in turbid biological media. Furthermore, we showcase their biological applications in cellular barcoding and microenvironmental sensing of a target protein and local electric field.
Highlights
Fluorescent optical probes have rapidly transformed our understanding of complex biological systems by providing specific information on biological targets in the natural living state
We observed that the simulated reflectance spectra are sinusoids when plotted against wavenumber, which is the inverse of wavelength (Fig. 1b, c, Supplementary Fig. 4 and Supplementary Note 2)
Using off-the-shelf reflectophores and the spectral reflectometry (SpeRe) imaging system, we showed that the measurement precision for physical size in our protocol was ±1.5 nm in an aqueous medium and ±3.0 nm in intracellular environments
Summary
Fluorescent optical probes have rapidly transformed our understanding of complex biological systems by providing specific information on biological targets in the natural living state Their utility is often limited by insufficient brightness, photostability, and multiplexing capacity. Reflectophores are orders-of-magnitudes brighter than conventional fluorophores and are free from photobleaching, enabling practically unlimited readout at high fidelity They offer high-degree multiplexing, encoded in their optical size, which can be readily decoded through interferometric detection with nanoscale accuracy, even in turbid biological media. Many strategies have been proposed to enable high-degree multiplexing, such as spectral unmixing[10], fluorescence lifetime[11], barcoded particles[12,13,14,15,16], and combinatoric labeling[17,18] These techniques have not been successfully adopted widely because they necessitate either complex fabrication processes or decoding optic systems. We report comprehensively on reflectophores from their theoretical background to their biological utilities in cellular barcoding and microenvironmental sensing
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