In summary, a tiny droplet of purified protein solution or cell suspension is placed on an EM grid and vitrified with the FEI Vitrobot. Subsequently, this sample is imaged using a transmission electron microscope like the Titan Krios, Tecnai Arctica or Talos and the resulting signal is detected using Falcon direct electron detection.
In Single Particle Analysis (SPA), automated image acquisition software EPU acquires the data which is processed using dedicated reconstruction software. The result is a high resolution 3D image of the viral particle or protein (complex).
Tomography is used to obtain ultra-structural 3D information from cells and cell organelles. The vitrified sample is imaged under various alpha tilts and the resulting data is subsequently processed to obtain the original 3D information.
By optimally linking high resolution SPA data into their natural biological context by using template matching software ARGOS, it is possible to localize the object of interest in its 3D context and to define the real orientation of the high resolution object. This adds an additional dimension which provides a better understanding of the functioning of molecular complexes, a cell or cell component.