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Correlative Cathodoluminescence Electron Microscopy: Immunolabeling Using Rare-Earth Element Doped Nanoparticles
Kerda Keevend1,2, Roman Krummenacher1, Egle Kungas1
1Laboratory for Particles Biology Interactions, Department Materials Meet Life, Swiss Federal Laboratories for Materials Science and Technology (Empa), Lerchenfeldstrasse 5, St. Gallen, CH-9014, Switzerland.
This study introduces cathodoluminescence (CL) immunotargeting, a novel electron microscopy technique using nanoparticle labels to visualize specific biomolecules and their structures at the nanoscale. This method enables precise protein localization within cellular ultrastructure, advancing molecular blueprint understanding.
Area of Science:
- Biophysics
- Nanotechnology
- Cell Biology
Background:
- Understanding biological systems requires nanoscale structure-function analysis.
- Electron microscopy (EM) offers high resolution but struggles with unambiguous molecular localization.
- A direct route to molecular blueprints necessitates EM methods for biomolecule localization within ultrastructure.
Purpose of the Study:
- To develop a novel EM approach for visualizing biomolecules within cellular ultrastructure.
- To harness cathodoluminescence (CL) for nanoscale molecular identification using spectral signatures.
- To demonstrate CL-enabled immunolabeling for protein localization in cancer cells.
Main Methods:
- Utilized cathodoluminescence (CL) microscopy, using an electron beam to generate optical emission at the nanoscale.
- Developed rare-earth element doped nanoparticle labels for specific biomolecule visualization.
- Employed folic acid decorated nanoparticles for targeted labeling of cell surface receptors.
Main Results:
- Demonstrated CL-enabled immunotargeting for nanoscale visualization of specific molecules within cellular ultrastructure.
- Successfully visualized characteristic receptor clustering on cancer cell surfaces using CL.
- Achieved specific molecular labeling based on spectral signatures rather than electron density.
Conclusions:
- CL immunotargeting provides nanoscale resolution for protein localization within cellular ultrastructure.
- This technique offers a direct pathway to understanding the molecular blueprints of biological systems.
- Paves the way for multiplexed protein immunolabeling in electron microscopy.
