skip to main content


This content will become publicly available on June 1, 2024

Title: Highly Sensitive Detection of Bacteria by Binder-Coupled Multifunctional Polymeric Dyes

Infectious diseases caused by pathogens are a health burden, but traditional pathogen identification methods are complex and time-consuming. In this work, we have developed well-defined, multifunctional copolymers with rhodamine B dye synthesized by atom transfer radical polymerization (ATRP) using fully oxygen-tolerant photoredox/copper dual catalysis. ATRP enabled the efficient synthesis of copolymers with multiple fluorescent dyes from a biotin-functionalized initiator. Biotinylated dye copolymers were conjugated to antibody (Ab) or cell-wall binding domain (CBD), resulting in a highly fluorescent polymeric dye-binder complex. We showed that the unique combination of multifunctional polymeric dyes and strain-specific Ab or CBD exhibited both enhanced fluorescence and target selectivity for bioimaging of Staphylococcus aureus by flow cytometry and confocal microscopy. The ATRP-derived polymeric dyes have the potential as biosensors for the detection of target DNA, protein, or bacteria, as well as bioimaging.

 
more » « less
Award ID(s):
2202747
NSF-PAR ID:
10495341
Author(s) / Creator(s):
; ; ; ; ; ;
Publisher / Repository:
MDPI
Date Published:
Journal Name:
Polymers
Volume:
15
Issue:
12
ISSN:
2073-4360
Page Range / eLocation ID:
2723
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
More Like this
  1. Abstract

    Small molecular NIR‐II dyes are highly desirable for various biomedical applications. However, NIR‐II probes are still limited due to the complex synthetic processes and inadequate availability of fluorescent core. Herein, the design and synthesis of three small molecular NIR‐II dyes are reported. These dyes can be excited at 850–915 nm and emitted at 1280–1290 nm with a large stokes shift (≈375 nm). Experimental and computational results indicate a 2:1 preferable host–guest assembly between the cucurbit[8]uril (CB) and dye molecules. Interestingly, the dyes when self‐assembled in presence of CB leads to the formation of nanocubes (≈200 nm) and exhibits marked enhancement in fluorescence emission intensity (Switch‐On). However, the addition of red carbon dots (rCDots, ≈10 nm) quenches the fluorescence of these host–guest complexes (Switch‐Off) providing flexibility in the user‐defined tuning of photoluminescence. The turn‐ON complex found to have comparable quantum yield to the commercially available near‐infrared fluorophore, IR‐26. The aqueous dispersibility, cellular and blood compatibility, and NIR‐II bioimaging capability of the inclusion complexes is also explored. Thus, a switchable fluorescence behavior, driven by host–guest complexation and supramolecular self‐assembly, is demonstrated here for three new NIR‐II dyes.

     
    more » « less
  2. Abstract

    The effect of vortex‐induced mechanical stresses on the fluorescent properties of dye‐containing poly(ethylene glycol)‐block‐poly(lactic acid) (PEG‐b‐PLA) block copolymer micelles has been investigated. PEG‐b‐PLA block copolymer micelles containing fluorescent dyes, 3,3′‐dioctadecyloxacarbocyanine perchlorate (DiO) and/or 1,1′‐dioctadecyl‐3,3,3′,3′‐tetramethylindocarbocyanine perchlorate (DiI), are prepared by a simple one‐step procedure that involves the self‐assembly of block copolymers and spontaneous incorporation of hydrophobic dyes into the core of the micelles. Upon vortexing, the micelle dispersion samples showed a decrease in fluorescence intensity in a rotational speed‐ and time‐dependent manner. The results demonstrated that the vortexing can alter the fluorescent properties of the dye‐containing PEG‐b‐PLA block copolymer micelle dispersion samples, suggesting the potential utility of block copolymer micelles as a mechanical stress‐responsive nanomaterial.

     
    more » « less
  3. Abstract

    Ultrabright fluorescent nanoparticles (NPs) hold great promise for demanding bioimaging applications. Recently, extremely bright molecular crystals of cationic fluorophores were obtained by hierarchical coassembly with cyanostar anion‐receptor complexes. These small‐molecule ionic isolation lattices (SMILES) ensure spatial and electronic isolation to prohibit aggregation quenching of dyes. We report a simple, one‐step supramolecular approach to formulate SMILES materials into NPs. Rhodamine‐based SMILES NPs stabilized by glycol amphiphiles show high fluorescence quantum yield (30 %) and brightness per volume (5000 M−1 cm−1/nm3) with 400 dye molecules packed into 16‐nm particles, corresponding to a particle absorption coefficient of 4×107 M−1 cm−1. UV excitation of the cyanostar component leads to higher brightness (>6000 M−1 cm−1/ nm3) by energy transfer to rhodamine emitters. Coated NPs stain cells and are thus promising for bioimaging.

     
    more » « less
  4. Abstract

    Ultrabright fluorescent nanoparticles (NPs) hold great promise for demanding bioimaging applications. Recently, extremely bright molecular crystals of cationic fluorophores were obtained by hierarchical coassembly with cyanostar anion‐receptor complexes. These small‐molecule ionic isolation lattices (SMILES) ensure spatial and electronic isolation to prohibit aggregation quenching of dyes. We report a simple, one‐step supramolecular approach to formulate SMILES materials into NPs. Rhodamine‐based SMILES NPs stabilized by glycol amphiphiles show high fluorescence quantum yield (30 %) and brightness per volume (5000 M−1 cm−1/nm3) with 400 dye molecules packed into 16‐nm particles, corresponding to a particle absorption coefficient of 4×107 M−1 cm−1. UV excitation of the cyanostar component leads to higher brightness (>6000 M−1 cm−1/ nm3) by energy transfer to rhodamine emitters. Coated NPs stain cells and are thus promising for bioimaging.

     
    more » « less
  5. null (Ed.)
    Fluorescent tagging is a popular method in biomedical research. Using multiple taggants of different but resolvable fluorescent spectra simultaneously (multiplexing), it is possible to obtain more comprehensive and faster information about various biochemical reactions and diseases, for example, in the method of flow cytometry. Here we report on a first demonstration of the synthesis of ultrabright fluorescent silica nanoporous nanoparticles (Star-dots), which have a large number of complex fluorescence spectra suitable for multiplexed applications. The spectra are obtained via simple physical mixing of different commercially available fluorescent dyes in a synthesizing bath. The resulting particles contain dye molecules encapsulated inside of cylindrical nanochannels of the silica matrix. The distance between the dye molecules is sufficiently small to attain Forster resonance energy transfer (FRET) coupling within a portion of the encapsulated dye molecules. As a result, one can have particles of multiple spectra that can be excited with just one wavelength. We show this for the mixing of five, three, and two dyes. Furthermore, the dyes can be mixed inside of particles in different proportions. This brings another dimension in the complexity of the obtained spectra and makes the number of different resolvable spectra practically unlimited. We demonstrate that the spectra obtained by different mixing of just two dyes inside of each particle can be easily distinguished by using a linear decomposition method. As a practical example, the errors of demultiplexing are measured when sets of a hundred particles are used for tagging. 
    more » « less