1. Page 1 Cryo-EM Services by Creative Biostructure

2. Page 2 Electron microscopyElectron microscopy (EM) has become an extremely popular method for the ultrastructural study of macromolecules, cells and tissues. With our in-house EM platform at Creative Biostructure, we are mainly focused on the use of cryo-EM technique to determine the structure of large macromolecules and their complexes with small ligands, proteins and nucleic acids. Other types of samples are also acceptable and can be handled with scanning electron microscopy (SEM) or transmission electron microscopy (TEM).scanning electron microscopy (SEM) transmission electron microscopy (TEM)

3. Page 3 (1) Sample preparation After protein purification, these samples will be treated by cryo-fixation. In this method, protein samples are placed on a specially treated EM grid consisting of tiny holes in a film supported by a metal frame. The grid is then plunged into liquid ethane to flash-freeze it, resulting in the protein samples being embedded in a thin layer of vitreous ice. Once the frozen hydrated grid is prepared, it is placed in the electron microscope and kept at approximately -180 K throughout the experiment.

4. Page 4 (2) EM imaging and data processing High-resolution EM images are obtained and data processed by our experienced scientists. We provide services that include: Data collection; Initial 3D model calculation; Beam-induced motion correction; Micrograph screening; Automatic particle picking and normalization; 2D, 3D classification and refinement.

5. Page 5 (3) Model building and refinement Using our high-performance computers and software packages, we are able to interpret EM maps and reconstruct them into three-dimensional structural models that satisfy principles of physics and stereochemistry. Our scientists are also very experienced with challenging targets such as membrane proteins by manual intervention to improve initial fit or even build de novo models. After an initial model is built, refinement is performed to maximize the agreement between the model and experimentally observed data by adjusting atomic coordinates, B factors and other parameters.

6. Page 6 Contact Creative Biostructure Mailing Address: 45-1 Ramsey Road Shirley, NY 11967, USA Email Address: contact@creative-biostructure.com Website: http://www.creative-biostructure.com/cryo-em- services_4.htmhttp://www.creative-biostructure.com/cryo-em- services_4.htm