Scanning Electron Microscopy

A scanning electron microscope uses an electron gun to generate a negatively charged electron beam, which is concentrated by electromagnetic lenses and deflected across the sample surface by scanning coils. The high-energy electron beam excites atoms in the sample, releasing low-energy secondary electrons that are detected to create a 3D image of the surface topography.

Electron-sample interactions produce secondary electrons from inelastic scattering, which reveal surface topography, and backscattered electrons from elastic scattering, which provide compositional information. The electron beam also generates characteristic X-rays that identify specific elements present in the sample, such as iron and nickel.

Secondary electrons are low-energy outer-shell electrons released by inelastic scattering, providing only topographical information from the sample surface. Backscattered electrons result from elastic scattering with the nucleus, maintaining their energy but changing direction, and reveal compositional differences based on atomic weight variations.

Conventional scanning electron microscopy provides magnification ranging from 20X to 30,000X with spatial resolution of 50 to 100 nanometers. This allows imaging of samples from approximately 1 centimeter down to 5 micrometers in width, making SEM suitable for bacteria, viruses, tissues, and larger specimens like insects.

When the incident electron beam undergoes inelastic collisions with electrons in discrete orbital shells of sample atoms, it excites those electrons to higher energy states. As the excited electrons return to lower energy levels, they emit characteristic X-rays with fixed wavelengths specific to each element, enabling elemental identification.

Backscattered electrons in SEM provide compositional information based on atomic weight differences. Biological samples can be studied to identify embedded or attached nanoparticles and nanostructures with heavier atomic weights, such as gold or iron, which produce varying contrast and allow distinction of sample composition.

Electromagnetic lenses concentrate the electron beam and reduce its diameter to achieve fine focus. Scanning coils then deflect this focused beam along the x- and y-axes, allowing systematic scanning of a rectangular area on the sample surface to build a complete topographical image.