Correlative cathodoluminescence electron microscopy bioimaging: towards single protein labelling with ultrastructural context

Abstract

<p>The understanding of living systems and their building blocks relies heavily on the assessment of structure-function relationships at the nanoscale. Ever since the development of the first optical microscope, the...</p>

Single Particle Cathodoluminescence Spectroscopy with Sub-20 nm, Electron-Stable Phosphors

“…We have demonstrated that these nanophosphors are sufficiently stable and bright enough to enable the collection of singleparticle CL spectra with average edge lengths of 15.2 ± 2.0 nm. In addition to addressing the urgent need to develop bright and stable cathodoluminescent nanophosphors 20 , we also explored the feasibility of our nanophosphor library for multiplexed spectral imaging. These experiments revealed that even at the single-particle level, band-pass filtered panchromatic CL images and CL spectra suffer from signal convolution by nearby nanophosphors.…”

“…Maturation and wide-spread adoption of EM-CL as an imaging technique still faces several distinctive challenges. 20 Of utmost importance is the development of bright and stable nanophosphors to enable single biomolecule labeling and localization. 20 In recent years, lanthanide-doped nanoparticles have emerged as promising candidates for cathodoluminescent nanophosphors.…”

“…20 Of utmost importance is the development of bright and stable nanophosphors to enable single biomolecule labeling and localization. 20 In recent years, lanthanide-doped nanoparticles have emerged as promising candidates for cathodoluminescent nanophosphors. Organic fluorophores or fluorescent proteins, commonly employed in 4 fluorescence microscopy, undergo radiolysis and rapidly degrade under electron irradiation.…”

“…Maturation and widespread adoption of EM-CL as an imaging technique still faces several distinctive challenges . Of utmost importance is the development of bright and stable nanophosphors to enable single biomolecule labeling and localization .…”

Luminescence properties of Gd(CO3)OH spherical particles with prospects for CL microscopic analysis and multi-color displays

“…13 Therefore, bright and stable phosphors with submicron to nanometer sizes are urgently needed for the development of CL microscopic analysis and FEDs, which requires single particle luminescence and positioning. 14 In recent years, high intensity electron beams with stable nanocrystals doped with different CL emitting elements have met the above requirements. 15,16 Among many nanostructures, gadolinium-based compounds have attracted much attention because of their applications in large flat panel displays, 17,18 magnetic resonance imaging contrast agents, 19,20 and so on.…”