Correlative cellular ptychography with functionalized nanoparticles at the Fe L-edge

Sci Rep. 2017 Jul 6;7(1):4757. doi: 10.1038/s41598-017-04784-5.

Abstract

Precise localization of nanoparticles within a cell is crucial to the understanding of cell-particle interactions and has broad applications in nanomedicine. Here, we report a proof-of-principle experiment for imaging individual functionalized nanoparticles within a mammalian cell by correlative microscopy. Using a chemically-fixed HeLa cell labeled with fluorescent core-shell nanoparticles as a model system, we implemented a graphene-oxide layer as a substrate to significantly reduce background scattering. We identified cellular features of interest by fluorescence microscopy, followed by scanning transmission X-ray tomography to localize the particles in 3D, and ptychographic coherent diffractive imaging of the fine features in the region at high resolution. By tuning the X-ray energy to the Fe L-edge, we demonstrated sensitive detection of nanoparticles composed of a 22 nm magnetic Fe3O4 core encased by a 25-nm-thick fluorescent silica (SiO2) shell. These fluorescent core-shell nanoparticles act as landmarks and offer clarity in a cellular context. Our correlative microscopy results confirmed a subset of particles to be fully internalized, and high-contrast ptychographic images showed two oxidation states of individual nanoparticles with a resolution of ~16.5 nm. The ability to precisely localize individual fluorescent nanoparticles within mammalian cells will expand our understanding of the structure/function relationships for functionalized nanoparticles.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Cells, Immobilized
  • Ferrosoferric Oxide / chemistry*
  • Fluorescence
  • Graphite / chemistry
  • HeLa Cells
  • Humans
  • Microscopy, Fluorescence / instrumentation
  • Microscopy, Fluorescence / methods*
  • Molecular Imaging / instrumentation
  • Molecular Imaging / methods*
  • Nanoparticles / chemistry*
  • Nanoparticles / ultrastructure
  • Oxidation-Reduction
  • Oxides / chemistry
  • Silicon Dioxide / chemistry
  • Tomography, X-Ray Computed / instrumentation
  • Tomography, X-Ray Computed / methods*

Substances

  • Oxides
  • Silicon Dioxide
  • Graphite
  • Ferrosoferric Oxide