Bright sub-20-nm cathodoluminescent nanoprobes for electron microscopy
Source abstract
Abstract
Electron microscopy has been instrumental in our understanding of complex biological systems. Although electron microscopy reveals cellular morphology with nanoscale resolution, it does not provide information on the location of different types of proteins. An electron-microscopy-based bioimaging technology capable of localizing individual proteins and resolving protein-protein interactions with respect to cellular ultrastructure would provide important insights into the molecular biology of a cell. Here, we synthesize small lanthanide-doped nanoparticles and measure the absolute photon emission rate of individual nanoparticles resulting from a given electron excitation flux (cathodoluminescence). Our results suggest that the optimization of nanoparticle composition, synthesis protocols and electron imaging conditions can lead to sub-20-nm nanolabels that would enable high signal-to-noise localization of individual biomolecules within a cellular context. In ensemble measurements, these labels exhibit narrow spectra of nine distinct colours, so the imaging of biomolecules in a multicolour electron microscopy modality may be possible.
Source: europepmc · Quality: candidate unverified · Source record ↗
Research context
OpenAlex topics: Advanced Electron Microscopy Techniques and Applications · Electron and X-Ray Spectroscopy Techniques · Advanced Fluorescence Microscopy Techniques
SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.
References (40)
Show source-provided references
1. Roth, J., Bendayan, M. & Orci, L. Ultrastructural localization of intracellular antigens by the use of protein A–gold complex. J. Histochem. Cytochem. 26, 1074–1081 (1978).10.1177/26.12.366014 ↗
2. Martell, J. D. et al. Engineered ascorbate peroxidase as a genetically encoded reporter for electron microscopy. Nat. Biotechnol. 30, 1143–1148 (2012).10.1038/nbt.2375 ↗
3. Lam, S. S. et al. Directed evolution of APEX2 for electron microscopy and proximity labeling. Nat. Methods 12, 51–54 (2015).10.1038/nmeth.3179 ↗
4. Adams, S. R. et al. Multicolor electron microscopy for simultaneous visualization of multiple molecular species. Cell Chem. Biol. 23, 1417–1427 (2016).10.1016/j.chembiol.2016.10.006 ↗
5. Sochacki, Ka, Shtengel, G., van Engelenburg, S. B., Hess, H. F. & Taraska, J. W. Correlative super-resolution fluorescence and metal-replica transmission electron microscopy. Nat. Methods 11, 305–308 (2014).10.1038/nmeth.2816 ↗
6. Carter, S. D. et al. Distinguishing signal from autofluorescence in cryogenic correlated light and electron microscopy of mammalian cells. J. Struct. Biol. 201, 15–25 (2018).10.1016/j.jsb.2017.10.009 ↗
7. Fisher, P. J., Wessels, W. S., Dietz, A. B. & Prendergast, F. G. Enhanced biological cathodoluminescence. Opt. Commun. 281, 1901–1908 (2008).10.1016/j.optcom.2007.04.069 ↗
8. Mahfoud, Z. et al. Cathodoluminescence in a scanning transmission electron microscope: a nanometer-scale counterpart of photoluminescence for the study of II–VI quantum dots. J. Phys. Chem. Lett. 4, 4090–4094 (2013).10.1021/jz402233x ↗
9. Zhang, H., Glenn, D. R., Schalek, R., Lichtman, J. W. & Walsworth, R. L. Efficiency of cathodoluminescence emission by nitrogen-vacancy color centers in nanodiamonds. Small 13, 1700543 (2017).10.1002/smll.201700543 ↗
10. Glenn, D. R. et al. Correlative light and electron microscopy using cathodoluminescence from nanoparticles with distinguishable colours. Sci. Rep. 2, 1–6 (2012).10.1038/srep00865 ↗
11. Niioka, H. et al. Multicolor cathodoluminescence microscopy for biological imaging with nanophosphors. Appl. Phys. Express 4, 2402 (2011).10.1143/APEX.4.112402 ↗
12. Cai, E. et al. Stable small quantum dots for synaptic receptor tracking on live neurons. Angew. Chem. Int. Ed. 53, 12484–12488 (2014).10.1002/anie.201405735 ↗
13. Yokota, S. Effect of particle size on labeling density for catalase in protein A–gold immunocytochemistry. J. Histochem. Cytochem. 36, 107–109 (1988).10.1177/36.1.3335766 ↗
14. Kijanka, M. et al. A novel immuno-gold labeling protocol for nanobody-based detection of HER2 in breast cancer cells using immuno-electron microscopy. J. Struct. Biol. 199, 1–11 (2017).10.1016/j.jsb.2017.05.008 ↗
15. Giepmans, B. N. G., Deerinck, T. J., Smarr, B. L., Jones, Y. Z. & Ellisman, M. H. Correlated light and electron microscopic imaging of multiple endogenous proteins using quantum dots. Nat. Methods 2, 743–749 (2005).10.1038/nmeth791 ↗
16. Bischak, C. G. et al. Cathodoluminescence-activated nanoimaging: noninvasive near-field optical microscopy in an electron microscope. Nano Lett. 15, 3383–3390 (2015).10.1021/acs.nanolett.5b00716 ↗
17. Feng, W., Zhu, X. & Li, F. Recent advances in the optimization and functionalization of upconversion nanomaterials for in vivo bioapplications. NPG Asia Mater. 5, e75–15 (2013).10.1038/am.2013.63 ↗
18. García De Abajo, F. J. Optical excitations in electron microscopy. Rev. Mod. Phys. 82, 209–275 (2010).10.1103/RevModPhys.82.209 ↗
19. Wang, F., Deng, R. & Liu, X. Preparation of core–shell NaGdF4 nanoparticles doped with luminescent lanthanide ions to be used as upconversion-based probes. Nat. Protoc. 9, 1634–1644 (2014).10.1038/nprot.2014.111 ↗
20. Liu, Q., Feng, W., Yang, T., Yi, T. & Li, F. Upconversion luminescence imaging of cells and small animals. Nat. Protoc. 8, 2033–2044 (2013).10.1038/nprot.2013.114 ↗
21. Li, Z. & Zhang, Y. An efficient and user-friendly method for the synthesis of hexagonal-phase NaYF4:Yb, Er/Tm nanocrystals with controllable shape and upconversion fluorescence. Nanotechnology 19, 345606 (2008).10.1088/0957-4484/19/34/345606 ↗
22. Ostrowski, A. D. et al. Controlled synthesis and single-particle imaging of bright, sub-10-nm lanthanide-doped upconverting nanocrystals. ACS Nano 6, 2686–2692 (2012).10.1021/nn3000737 ↗
23. Dong, A. et al. A generalized ligand-exchange strategy enabling sequential surface functionalization of colloidal nanocrystals. J. Am. Chem. Soc. 133, 998–1006 (2011).10.1021/ja108948z ↗
24. Gargas, D. J. et al. Engineering bright sub-10-nm upconverting nanocrystals for single-molecule imaging. Nat. Nanotechnol. 9, 300–305 (2014).10.1038/nnano.2014.29 ↗
25. Feng, W., Sun, L. D., Zhang, Y. W. & Yan, C. H. Solid-to-hollow single-particle manipulation of a self-assembled luminescent NaYF4:Yb,Er nanocrystal monolayer by electron-beam lithography. Small 5, 2057–2060 (2009).10.1002/smll.200900404 ↗
26. Judd, B. R. Optical absorption intensities of rare-earth ions. Phys. Rev. 127, 750–761 (1962).10.1103/PhysRev.127.750 ↗
27. Ofelt, G. S. Intensities of crystal spectra of rare‐earth ions. J. Chem. Phys. 37, 511–520 (1962).10.1063/1.1701366 ↗
28. Chan, E. M. Combinatorial approaches for developing upconverting nanomaterials: high-throughput screening, modeling, and applications. Chem. Soc. Rev. 44, 1653–1679 (2015).10.1039/C4CS00205A ↗
29. Fischer, S., Johnson, N. J. J., Pichaandi, J., Goldschmidt, J. C. & van Veggel, F. C. J. M. Upconverting core–shell nanocrystals with high quantum yield under low irradiance: on the role of isotropic and thick shells. J. Appl. Phys. 118, 193105 (2015).10.1063/1.4936119 ↗
30. Ma, C. et al. Optimal sensitizer concentration in single upconversion nanocrystals. Nano Lett. 17, 2858–2864 (2017).10.1021/acs.nanolett.6b05331 ↗
31. Lu, Y. et al. Tunable lifetime multiplexing using luminescent nanocrystals. Nat. Photon. 8, 32–36 (2013).10.1038/nphoton.2013.322 ↗
32. Nawa, Y. et al. Dynamic autofluorescence imaging of intracellular components inside living cells using direct electron beam excitation. Biomed. Opt. Express 5, 378–386 (2014).10.1364/BOE.5.000378 ↗
33. Xu, C. S. et al. Enhanced FIB-SEM systems for large-volume 3D imaging. eLife 3, 1–36 (2017).
34. Zhang, Z., Kenny, S. J., Hauser, M., Li, W. & Xu, K. Ultrahigh-throughput resolved super-resolution microscopy. Nat. Methods 12, 935–938 (2015).10.1038/nmeth.3528 ↗
35. DaCosta, M. V., Doughan, S., Han, Y. & Krull, U. J. Lanthanide upconversion nanoparticles and applications in bioassays and bioimaging: a review. Anal. Chim. Acta 832, 1–33 (2014).10.1016/j.aca.2014.04.030 ↗
36. Denk, W. & Horstmann, H. Serial block-face scanning electron microscopy to reconstruct three-dimensional tissue nanostructure. PLoS Biol. 2, e329 (2004).
37. Hayworth, K. J. et al. Ultrastructurally smooth thick partitioning and volume stitching for large-scale connectomics. Nat. Methods 12, 319–322 (2015).10.1038/nmeth.3292 ↗
38. Kasthuri, N. et al. Saturated reconstruction of a volume of neocortex. Cell 162, 648–661 (2015).10.1016/j.cell.2015.06.054 ↗
39. Stephenson, R. S. et al. High resolution 3-dimensional imaging of the human cardiac conduction system from microanatomy to mathematical modeling. Sci. Rep. 7, 7188 (2017).10.1038/s41598-017-07694-8 ↗
40. Angelo, M. et al. Multiplexed ion beam imaging of human breast tumors. Nat. Med. 20, 436–442 (2014).10.1038/nm.3488 ↗