Cryogenic Electron Microscopy (Cryo-EM) is a technique that allows the study of specimens at cryogenic temperatures, preserving them in their native state without staining or fixing. This technique is particularly powerful for exploring complex biological structures at high resolution. Cryo-electron tomography (cryo-ET) creates 3D reconstructions from tilted 2D images, while single-particle analysis (SPA) achieves high resolution through extensive statistical analysis of multiple copies of the same molecule.

Correlative Light-Electron Microscopy (CLEM) is a cutting-edge technique that combines optical microscopy (usually fluorescence) with electron microscopy (EM), enabling detailed imaging across different scales. EM provides high-resolution information down to the nanoscale, while the fluorescence microscope highlights regions of interest, provides molecular information, and allows for live imaging. The Netherlands leads in CLEM innovation, with significant contributions to efficient, high-precision methods and integrated microscope development. The Netherlands is a key player in the ESFRI EuroBioImaging initiative, with the Dutch CLEM community serving as the primary entry for CLEM. As part of NEMI, strategic investments in high-precision section CLEM, live cell CLEM, and cryo-CLEM aim to enhance correlation speed and accuracy, and to drive the implementation of cryo-ET by defining regions of interest for imaging.

Multimodal Electron Microscopy (EM) combines high-resolution EM with mass spectrometry (MS) to visualize the spatial distribution of molecules by their masses, providing unprecedented insights into the chemistry of complex specimens. Combining high-resolution EM and high-resolution MS in a single instrument for cryo-preserved specimens enables the study of (bio)chemistry in three dimensions of complex volumes.

Multibeam Electron Microscopy (EM) is designed to significantly reduce acquisition time by using multiple electron beams simultaneously, enabling high-contrast imaging of large areas and volumes. This technology is crucial for capturing detailed images of complex biological specimens, such as entire organs or tumor cells.

• High-Resolution Large-Area Imaging: Stitching many images together to cover millimeter-sized areas or imaging multiple slices to create 3D images.
• Google-Map Type Viewing: The UMCG has developed a setup that stitches many images into super-images for detailed, large-scale viewing.
• Multibeam SEM Development: TUD has developed a unique multibeam SEM with 196 beams and transmission detection, specifically for high-contrast imaging of thin biological specimens.

High-resolution Electron Microscopy (EM) in materials science is crucial for advancing our understanding and development of new materials, especially in sustainability and future energy resources. Dutch researchers are renowned for their work in in-situ EM, which allows real-time imaging of transformations and chemical processes at atomic resolution. This capability is essential for studying dynamic changes in materials under various conditions.