RESUMO
Extracellular vesicles (EVs) have great potential as biomarkers since their composition and concentration in biofluids are disease state dependent and their cargo can contain disease-related information. Large tumor-derived EVs (tdEVs, >1 µm) in blood from cancer patients are associated with poor outcome, and changes in their number can be used to monitor therapy effectiveness. Whereas, small tumor-derived EVs (<1 µm) are likely to outnumber their larger counterparts, thereby offering better statistical significance, identification and quantification of small tdEVs are more challenging. In the blood of cancer patients, a subpopulation of EVs originate from tumor cells, but these EVs are outnumbered by non-EV particles and EVs from other origin. In the Dutch NWO Perspectief Cancer-ID program, we developed and evaluated detection and characterization techniques to distinguish EVs from non-EV particles and other EVs. Despite low signal amplitudes, we identified characteristics of these small tdEVs that may enable the enumeration of small tdEVs and extract relevant information. The insights obtained from Cancer-ID can help to explore the full potential of tdEVs in the clinic.
RESUMO
Correlative and integrated scanning electron microscopy (SEM) and Raman micro-spectroscopy is presented that enables the characterization and identification of different cancer and non-cancer cells through SEM-Raman image cytometry. The hybrid microscopy system enables the acquisition of high resolution SEM images of uncoated cells and the spatial correlation with chemical information as obtained from Raman micro-spectroscopic imaging. A sample preparation protocol and a workflow are presented that are compatible with the demands of hybrid SEM-Raman microscopy. Stainless steel cell substrates were used that are both conductive and give a low optical response in Raman scattering. Correlative and integrated SEM-Raman micro-spectroscopy is illustrated with cells from blood and cells from a SKBR-3 breast cancer cell line.