Single particle cryo-electron microscopy (cryo-EM) is an emerging powerful tool for structural studies of macromolecular assemblies. Although less concentrated and smaller amounts of samples are required for single particle cryo-EM compared to X-ray crystallography, it remains challenging to study specimens that are low-abundance, low-yield, or short-lived. The recent development of affinity grid techniques holds great promise to tackle these challenging samples by combining the sample purification and freezing on TEM grids steps in cryo-EM grid preparation into a single step, revolutionize the grid preparation of cryo-EM, and extend single particle cryo-EM to a routine structural biology tool to characterize structures of a broad spectrum of macromolecules.
In my PhD study, I have established a new design of the affinity cryo-EM approach, cryo-SPIEM that applies a traditional pathogen diagnosis tool Solid Phase Immune Electron Microscopy (SPIEM) to the single particle cryo-EM method, and also systematically explored the applications of affinity EM approaches and the potentials of affinity cryo-EM approaches for near-atomic single particle 3-D reconstruction. The cryo-SPIEM approach provides an alternative, largely simplified and easier to use affinity grid that directly works with most native macromolecular complexes with established antibodies, and enables cryo-EM studies of native samples directly from cell cultures. The application of the affinity cryo-EM approach for high-resolution cryo-EM has been demonstrated successfully by solving a 6.3 Å structure of Tulane virus using the polylysine-based affinity grid, and a 2.6 Å structure of Tulane virus using the antibody-based affinity grid with a sample of low concentrations that defies standard cryo-EM study. Moreover, we have applied the affinity grid technique to investigate the interaction between Tulane virus and its cellular receptor Histo-blood group antigens, which revealed the potential roles of HBGA receptors in mediating genome release.
","abstract_html":"<p>Single particle cryo-electron microscopy (cryo-EM) is an emerging powerful tool for structural studies of macromolecular assemblies. Although less concentrated and smaller amounts of samples are required for single particle cryo-EM compared to X-ray crystallography, it remains challenging to study specimens that are low-abundance, low-yield, or short-lived. The recent development of affinity grid techniques holds great promise to tackle these challenging samples by combining the sample purification and freezing on TEM grids steps in cryo-EM grid preparation into a single step, revolutionize the grid preparation of cryo-EM, and extend single particle cryo-EM to a routine structural biology tool to characterize structures of a broad spectrum of macromolecules.</p> <p>In my PhD study, I have established a new design of the affinity cryo-EM approach, cryo-SPIEM that applies a traditional pathogen diagnosis tool Solid Phase Immune Electron Microscopy (SPIEM) to the single particle cryo-EM method, and also systematically explored the applications of affinity EM approaches and the potentials of affinity cryo-EM approaches for near-atomic single particle 3-D reconstruction. The cryo-SPIEM approach provides an alternative, largely simplified and easier to use affinity grid that directly works with most native macromolecular complexes with established antibodies, and enables cryo-EM studies of native samples directly from cell cultures. The application of the affinity cryo-EM approach for high-resolution cryo-EM has been demonstrated successfully by solving a 6.3 Å structure of Tulane virus using the polylysine-based affinity grid, and a 2.6 Å structure of Tulane virus using the antibody-based affinity grid with a sample of low concentrations that defies standard cryo-EM study. Moreover, we have applied the affinity grid technique to investigate the interaction between Tulane virus and its cellular receptor Histo-blood group antigens, which revealed the potential roles of HBGA receptors in mediating genome release.</p>","abstract_has_math":false,"creators":["Yu, Guimei"],"institution":null,"degree_name":"Doctor of Philosophy (PhD)","degree_level":"Dissertation","degree_discipline":"Biological Science","degree_department":null,"school":null,"contributors":["Wen Jiang","Richard J. Kuhn","Jason Lanman","Chiwook Park"],"advisors":[],"committee_chairs":[],"committee_members":[],"year":2016,"date_issued":"2016-05-01T07:00:00Z","date_published":"2016-05-01T07:00:00Z","updated_at":"2026-07-24T03:53:55Z","subjects":["Biophysics","Microbiology","Molecular Biology"],"languages":[],"rights":[],"rights_urls":[],"identifier_entries":[]},"links":{"outbound_url":"https://docs.lib.purdue.edu/open_access_dissertations/1041","outbound_label":"Repository record","outbound_source":"dc:identifier"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor","label":"Contributor","values":["Wen Jiang","Richard J. Kuhn","Jason Lanman","Chiwook Park"]},{"key":"dc:creator","label":"Author","values":["Yu, Guimei"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"thesis:degree_discipline","label":"Discipline","values":["Biological Science"]},{"key":"thesis:degree_level","label":"Degree Level","values":["Dissertation"]},{"key":"thesis:degree_name","label":"Degree Name","values":["Doctor of Philosophy (PhD)"]}]},{"id":"subjects_keywords","label":"Subjects and Keywords","entries":[{"key":"dc:subject","label":"Dc Subject","values":["Biophysics","Microbiology","Molecular Biology"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier","label":"Identifier","values":["https://docs.lib.purdue.edu/open_access_dissertations/1041"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Single particle cryo-electron microscopy (cryo-EM) is an emerging powerful tool for structural studies of macromolecular assemblies. Although less concentrated and smaller amounts of samples are required for single particle cryo-EM compared to X-ray crystallography, it remains challenging to study specimens that are low-abundance, low-yield, or short-lived. The recent development of affinity grid techniques holds great promise to tackle these challenging samples by combining the sample purification and freezing on TEM grids steps in cryo-EM grid preparation into a single step, revolutionize the grid preparation of cryo-EM, and extend single particle cryo-EM to a routine structural biology tool to characterize structures of a broad spectrum of macromolecules.
In my PhD study, I have established a new design of the affinity cryo-EM approach, cryo-SPIEM that applies a traditional pathogen diagnosis tool Solid Phase Immune Electron Microscopy (SPIEM) to the single particle cryo-EM method, and also systematically explored the applications of affinity EM approaches and the potentials of affinity cryo-EM approaches for near-atomic single particle 3-D reconstruction. The cryo-SPIEM approach provides an alternative, largely simplified and easier to use affinity grid that directly works with most native macromolecular complexes with established antibodies, and enables cryo-EM studies of native samples directly from cell cultures. The application of the affinity cryo-EM approach for high-resolution cryo-EM has been demonstrated successfully by solving a 6.3 Å structure of Tulane virus using the polylysine-based affinity grid, and a 2.6 Å structure of Tulane virus using the antibody-based affinity grid with a sample of low concentrations that defies standard cryo-EM study. Moreover, we have applied the affinity grid technique to investigate the interaction between Tulane virus and its cellular receptor Histo-blood group antigens, which revealed the potential roles of HBGA receptors in mediating genome release.
"]},{"key":"dc:title","label":"Title","values":["Affinity cryo-electron microscopy: Methods development and applications"]}]}],"canonical_facts":{"dc:contributor":["Wen Jiang","Richard J. Kuhn","Jason Lanman","Chiwook Park"],"dc:creator":["Yu, Guimei"],"dc:description.abstract":["Single particle cryo-electron microscopy (cryo-EM) is an emerging powerful tool for structural studies of macromolecular assemblies. Although less concentrated and smaller amounts of samples are required for single particle cryo-EM compared to X-ray crystallography, it remains challenging to study specimens that are low-abundance, low-yield, or short-lived. The recent development of affinity grid techniques holds great promise to tackle these challenging samples by combining the sample purification and freezing on TEM grids steps in cryo-EM grid preparation into a single step, revolutionize the grid preparation of cryo-EM, and extend single particle cryo-EM to a routine structural biology tool to characterize structures of a broad spectrum of macromolecules.
In my PhD study, I have established a new design of the affinity cryo-EM approach, cryo-SPIEM that applies a traditional pathogen diagnosis tool Solid Phase Immune Electron Microscopy (SPIEM) to the single particle cryo-EM method, and also systematically explored the applications of affinity EM approaches and the potentials of affinity cryo-EM approaches for near-atomic single particle 3-D reconstruction. The cryo-SPIEM approach provides an alternative, largely simplified and easier to use affinity grid that directly works with most native macromolecular complexes with established antibodies, and enables cryo-EM studies of native samples directly from cell cultures. The application of the affinity cryo-EM approach for high-resolution cryo-EM has been demonstrated successfully by solving a 6.3 Å structure of Tulane virus using the polylysine-based affinity grid, and a 2.6 Å structure of Tulane virus using the antibody-based affinity grid with a sample of low concentrations that defies standard cryo-EM study. Moreover, we have applied the affinity grid technique to investigate the interaction between Tulane virus and its cellular receptor Histo-blood group antigens, which revealed the potential roles of HBGA receptors in mediating genome release.
"],"dc:identifier":["https://docs.lib.purdue.edu/open_access_dissertations/1041"],"dc:subject":["Biophysics","Microbiology","Molecular Biology"],"dc:title":["Affinity cryo-electron microscopy: Methods development and applications"],"thesis:degree_discipline":["Biological Science"],"thesis:degree_level":["Dissertation"],"thesis:degree_name":["Doctor of Philosophy (PhD)"]},"updated_at":"2026-07-24T03:53:55Z"}