Opportunities and challenges in liquid cell electron microscopy

Advances in seeing small things Electron microscopes, particularly those with aberration correction, can view materials at the subnanometer scale. Additional improvements make it possible to obtain images at lower electron doses, thus minimizing the damage to the sample. However, for a number of materials, particularly those of biological origin, samples need to be imaged in solution. Ross reviews recent advances that have made it possible to do liquid cell electron microscopy, which opens up the possibility of studying problems such as the changes inside a battery during operation, the growth of crystals from solution, or biological molecules in their native state. Science, this issue p. 10.1126/science.aaa9886 Structured Abstract BACKGROUNDTransmission electron microscopy offers structural and compositional information with atomic resolution, but its use is restricted to thin, solid samples. Liquid samples, particularly those involving water, have been challenging because of the need to form a thin liquid layer that is stable within the microscope vacuum. Liquid cell electron microscopy is a developing technique that allows us to apply the powerful capabilities of the electron microscope to the imaging and analysis of liquid specimens. We can examine liquid-based processes in materials science and physics that are traditionally inaccessible to electron microscopy, and image biological structures at high resolution without the need for freezing or drying. The changes that occur inside batteries during operation, the attachment of atoms during the self-assembly of nanocrystals, and the structures of biological materials in liquid water are examples in which a microscopic view is providing unique insights. ADVANCESThe difficulty of imaging water and other liquids was recognized from the earliest times in the development of transmission electron microscopy. Achieving a practical solution, however, required the use of modern microfabrication techniques to build liquid cells with thin but strong windows. Usually made of silicon nitride on a silicon support, these liquid cells perform two jobs: They separate the liquid from the microscope vacuum while also confining it into a layer that is thin enough for imaging with transmitted electrons. Additional functionality such as liquid flow, electrodes, or heating can be incorporated in the liquid cell. The first experiments to make use of modern liquid cells provided information on electrochemical deposition, nanomaterials synthesis, diffusion in liquids, and the structure of biological assemblies. Materials and processes now under study include corrosion, biomolecular structure, bubble dynamics, radiation effects, and biomineralization. New window materials such as graphene can improve resolution, and elemental analysis is possible by measuring energy loss or x-ray signals. Advances in electron optics and detectors, and the correlation of liquid cell microscopy data with probes such as fluorescence, have increased the range of information available from the sample. Because the equipment is not too expensive and works in existing electron microscopes, liquid cell microscopy programs have developed around the world. OUTLOOKLiquid cell electron microscopy is well positioned to explore new frontiers in electrochemistry and catalysis, nanomaterial growth, fluid physics, diffusion, radiation physics, geological and environmental processes involving clays and aerosols, complex biomaterials and polymers, and biological functions in aqueous environments. Continuing improvements in equipment and technique will allow materials and processes to be studied under different stimuli—for example, in extreme temperatures, during gas/liquid mixing, or in magnetic or electric fields. Correlative approaches that combine liquid cell electron microscopy with light microscope or synchrotron data promise a deeper study of chemical, electrochemical, and photochemical reactions; analytical electron microscopy will provide details of composition and chemical bonding in water; high-speed and aberration-corrected imaging extend the scales of the phenomena that can be examined. As liquid cell microscopy becomes more capable and quantitative, it promises the potential to extend into new areas, adopt advanced imaging modes such as holography, and perhaps even solve grand challenge problems such as the structure of the electrochemical double layer or molecular movements during biological processes. Download high-res image Open in new tab Download Powerpoint Schematic diagram of a liquid cell for the transmission electron microscope and its application for imaging phenomena in materials science, life science, and physics.The liquid cell is made from two vacuum-tight electron transparent membranes. In this diagram the membranes are made of silicon nitride (blue) on a silicon support (gray), although other materials are possible. A spacer layer (not shown) keeps the membranes at a controlled separation of about 100 nm to 1 mm. The cell is filled with the liquid of interest, and the liquid may be flowed using an external pump (not shown). The electron beam (pink) passes through the membranes and liquid to allow recording of images, movies, or spectroscopic data for compositional analysis. Several possible experiments are illustrated: growth of nanocrystals in solution, nucleation and growth of bubbles, imaging biological structures such as whole cells or viruses in liquid water, and imaging electrochemical processes at an electrode (yellow) that is built into the liquid cell. The dimensions of the electron beam and the nanoscale objects are exaggerated for clarity. Transmission electron microscopy offers structural and compositional information with atomic resolution, but its use is restricted to thin, solid samples. Liquid samples, particularly those involving water, have been challenging because of the need to form a thin liquid layer that is stable within the microscope vacuum. Liquid cell electron microscopy is a developing technique that allows us to apply the powerful capabilities of the electron microscope to imaging and analysis of liquid specimens. We describe its impact in materials science and biology. We discuss how its applications have expanded via improvements in equipment and experimental techniques, enabling new capabilities and stimuli for samples in liquids, and offering the potential to solve grand challenge problems.