Recent technological developments in live cell microscopy and CLEM (correlative light and electron microscopy) are providing unique tools to characterize complex biological systems. We are developing new methods for molecular mapping in cryo-electron microscopy and tomography.
CLEM methods consist in the selection of individual live cells with interesting features for subsequent analysis at higher resolution. Ultrastructural studies in two and three dimensions can be done with a variety of electron microscopy techniques. We can now visualize cells under more native conditions thanks to developments made in specimen preparation at low temperatures (cryo-microscopy). Our lab has developed new probes for CLEM and (cryo)-electron tomography. The method, that uses metal-binding proteins as clonable tags, is compatible with a variety of sample processing techniques. Its applicability and strength is demonstrated in several studies which unveiled virus-induced cell structures not seen before.
Risco, Viruses 2021; Fernández de Castro et al.,Bioprotocol 2021; García-Serradilla & Risco. Virus Research 2021; Sachse et al., Advances in Virus Research 2019; Sachse et al., Methods in Molecular Biology 2018; Fernández de Castro & Risco. Bio-Protocol 2018; Fernández de Castro et al., Nature Communications 2017; Fernández de Castro et al., Journal of Cell Science 2017; Fernández de Castro et al., Methods in Cell Bioloy 2014; Barajas, Fernández de Castro et al., PLoS Pathogens 2014.