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DNA-Functionalized Nanoparticles for Multicolor Cathodoluminescence Imaging.

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This study introduces DNA-functionalized lanthanide nanoparticles (LNPs) for cathodoluminescence (CL) microscopy. These hydrophilic probes enable simultaneous visualization of proteins and cellular ultrastructure in biological imaging.

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Area of Science:

  • Biophysics
  • Nanotechnology
  • Microscopy

Background:

  • Cathodoluminescence (CL) microscopy offers potential for simultaneous visualization of proteins and cellular ultrastructure.
  • Lanthanide nanoparticles (LNPs) are suitable probes due to electron beam stability and distinct emission spectra for multiplexed detection.
  • The hydrophobic nature of LNPs necessitates surface functionalization for biological applications and electron microscopy (EM) compatibility.

Purpose of the Study:

  • To develop hydrophilic cathodoluminescent probes for biological imaging.
  • To enable simultaneous visualization of proteins and cellular ultrastructure using CL microscopy.
  • To demonstrate multicolor CL imaging with functionalized LNPs.

Main Methods:

  • Utilized a DNA-based ligand exchange strategy to functionalize LNPs, rendering them hydrophilic.
  • Characterized CL emission of DNA-functionalized LNPs after aqueous transfer and EM preparation steps (osmium tetroxide staining, HMDS, critical point drying).
  • Demonstrated multicolor CL imaging on mammalian cells using spectrally distinct, DNA-functionalized LNPs.

Main Results:

  • DNA-functionalized LNPs maintained CL emission throughout aqueous transfer and EM preparation.
  • Tested EM preparation methods included osmium tetroxide staining and drying protocols (HMDS, critical point drying).
  • Successfully achieved multicolor CL imaging of LNPs on mammalian cells.

Conclusions:

  • DNA-functionalization provides a viable method to create hydrophilic LNPs for biological CL imaging.
  • Functionalized LNPs are compatible with standard EM sample preparation techniques.
  • This approach enables simultaneous ultrastructural and multiplexed protein visualization in biological samples.