Molecularly imprinted polymers can be used as "plastic antibodies" for cell and tissue imaging, as demonstrated using hyaluronan on cell surfaces as a model target. Fluorescent nanoparticles binding a hyaluronan substructure, glucuronic acid, are used to image fixated and living cells and tissues. Plastic antibodies can be tailored to specific targets and easily labeled, and are physically and chemically stable.
Advanced tools for cell imaging are of great interest for the detection, localization, and quantification of molecular biomarkers of cancer or infection. We describe a novel photopolymerization method to coat quantum dots (QDs) with polymer shells, in particular, molecularly imprinted polymers (MIPs), by using the visible light emitted from QDs excited by UV light. Fluorescent core-shell particles specifically recognizing glucuronic acid (GlcA) or N-acetylneuraminic acid (NANA) were prepared. Simultaneous multiplexed labeling of human keratinocytes with green QDs conjugated with MIP-GlcA and red QDs conjugated with MIP-NANA was demonstrated by fluorescence imaging. The specificity of binding was verified with a non-imprinted control polymer and by enzymatic cleavage of the terminal GlcA and NANA moieties. The coating strategy is potentially a generic method for the functionalization of QDs to address a much wider range of biocompatibility and biorecognition issues.
Advanced tools for cell imaging are of great interest for the detection, localization, and quantification of molecular biomarkers of cancer or infection. We describe anovel photopolymerization method to coat quantum dots (QDs) with polymer shells,i np articular,m olecularly imprinted polymers (MIPs), by using the visible light emitted from QDs excited by UV light. Fluorescent core-shell particles specifically recognizing glucuronic acid (GlcA) or N-acetylneuraminic acid (NANA) were prepared. Simultaneous multiplexed labeling of human keratinocytes with green QDs conjugated with MIP-GlcA and red QDs conjugated with MIP-NANAwas demonstrated by fluorescence imaging. The specificity of binding was verified with an on-imprinted control polymer and by enzymatic cleavage of the terminal GlcA and NANA moieties.The coating strategy is potentially ag eneric method for the functionalization of QDs to address am uchw ider range of biocompatibility and biorecognition issues.Supportinginformation for this article, includinge xperimental details (reagents and materials, synthesis and characterization of MIP-QDs, equilibrium and competitive binding assays, TEM and DLS analysis, 1 HNMR studies of the AB-template complex, and cell and tissue staining and imaging), can be found under: http://dx.
obtained by colloidal templating, [ 22,23 ] since they allow the rapid fabrication of macroscale fi lms with homogeneous optical properties over a large surface area and also offer fl exibility to produce virtual optical elements and other types of indicators (bar graphs, images etc.) that are useful in visual sensing. The multiscale structuring of MI-SHRH offered by i) the macroscale of the polymer fi lm, ii) the microscale of the polymer pores to provide access to the binding sites, iii) the nanoscale of holographic fringe spacing, and iv) the molecular imprints, make MI-SHRH completely innovative hierarchicallystructured functional materials having the potential to give rise to inexpensive, mass-producible label-free sensing devices.
Advanced tools for cell imaging are of particular interest as they can detect, localize and quantify molecular targets like abnormal glycosylation sites that are biomarkers of cancer and infection. Targeting these biomarkers is often challenging due to a lack of receptor materials. Molecularly imprinted polymers (MIPs) are promising artificial receptors; they can be tailored to bind targets specifically, be labeled easily, and are physically and chemically stable. Herein, we demonstrate the application of MIPs as artificial antibodies for selective labeling and imaging of cellular targets, on the example of hyaluronan and sialylation moieties on fixated human skin cells and tissues. Thus, fluorescently labeled MIP nanoparticles templated with glucuronic acid (MIPGlcA) and N-acetylneuraminic acid (MIPNANA) are respectively applied. Two different fluorescent probes are used: (1) MIPGlcA particles, ~400 nm in size are labeled with the dye rhodamine that target the extracellular hyaluronan on cells and tissue specimens and (2) MIP-coated InP/ZnS quantum dots (QDs) of two different colors, ~125 nm in size that target the extracellular and intracellular hyaluronan and sialylation sites. Green and red emitting QDs are functionalized with MIPGlcA and MIPNANA respectively, enabling multiplexed cell imaging. This is a general approach that can also be adapted to other target molecules on and in cells.
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