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Acoustofluidic cryo-EM enables in situ particle manipulation for uniform, high-quality cryo-EM specimens.

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Title: Acoustofluidic cryo-EM enables in situ particle manipulation for uniform, high-quality cryo-EM specimens.
Authors: Yang K; Thomas Lord Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA., Peele W; Genome Integrity and Structural Biology Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Department of Health and Human Services, Research Triangle Park, Durham, NC 27709, USA., Liu P; Thomas Lord Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA., Xia J; Thomas Lord Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA., Dandey V; Genome Integrity and Structural Biology Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Department of Health and Human Services, Research Triangle Park, Durham, NC 27709, USA., Zhong R; Thomas Lord Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA., Jin K; Thomas Lord Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA., Sharma K; Genome Integrity and Structural Biology Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Department of Health and Human Services, Research Triangle Park, Durham, NC 27709, USA., Watson AJ; Genome Integrity and Structural Biology Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Department of Health and Human Services, Research Triangle Park, Durham, NC 27709, USA.; Department of Biochemistry, Duke University School of Medicine, Durham, NC 27710, USA., Yang S; Thomas Lord Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA., Chen Y; Thomas Lord Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA., Agahtehrani A; Thomas Lord Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA., Lan SN; Thomas Lord Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA., Zhang P; Thomas Lord Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA., Tian Z; Thomas Lord Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA., Liu M; Thomas Lord Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA., Viverette E; Genome Integrity and Structural Biology Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Department of Health and Human Services, Research Triangle Park, Durham, NC 27709, USA., Xu X; Thomas Lord Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA., Wu Q; Thomas Lord Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA., Borgnia MJ; Genome Integrity and Structural Biology Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Department of Health and Human Services, Research Triangle Park, Durham, NC 27709, USA., Lee LP; Harvard Medical School, Harvard University; and Renal Division and Division of Engineering in Medicine, Department of Medicine, Brigham and Women's Hospital, Boston, MA 02115, USA.; Department of Bioengineering, University of California, Berkeley, Berkeley, CA 94720, USA.; Department of Electrical Engineering and Computer Science, University of California, Berkeley, Berkeley, CA 94720, USA.; Department of Biophysics, Institute of Quantum Biophysics, Sungkyunkwan University, Suwon, Korea.; Department of Chemistry and Nanoscience, Ewha Womans University, Seoul, Korea., Huang TJ; Thomas Lord Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA.
Source: Science advances [Sci Adv] 2026 Sep 25; Vol. 12 (39), pp. eaeh5900. Date of Electronic Publication: 2026 Sep 23.
Publication Type: Journal Article
Language: English
Journal Info: Publisher: American Association for the Advancement of Science Country of Publication: United States NLM ID: 101653440 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 2375-2548 (Electronic) Linking ISSN: 23752548 NLM ISO Abbreviation: Sci Adv Subsets: MEDLINE
Imprint Name(s): Original Publication: Washington, DC : American Association for the Advancement of Science, [2015]-
MeSH Terms: Cryoelectron Microscopy*/methods , Cryoelectron Microscopy*/instrumentation , Specimen Handling*/methods , Acoustics*, Apoferritins/chemistry ; Liposomes/chemistry ; Liposomes/ultrastructure
Abstract: Cryo-electron microscopy (cryo-EM) has revolutionized structural biology by enabling imaging of macromolecular complexes at near-atomic resolution, but its success depends critically on specimen quality. Standard plunge freezing often produces nonuniform particle distributions, resulting in poor particle statistics and reduced imaging quality, particularly for vesicular structures and low-concentration, labile macromolecular complexes. Here, we introduce acoustofluidic cryo-EM (ACE), a noncontact specimen-preparation method that integrates acoustofluidic manipulation into a conventional plunge-freezing workflow. ACE enables localized nanoscale particle manipulation by mechanically coupling acoustic excitation into the cryo-EM grid, where forced vibration generates localized acoustic streaming through viscous boundary-layer effects. Guided by this physical mechanism, validation using liposomes and apoferritin demonstrates that ACE increases particle density by up to fourfold while preserving structural integrity and improving imaging quality. This strategy provides a broadly applicable approach for enhancing cryo-EM specimen quality, improving imaging efficiency, and facilitating high-resolution structural analysis.
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Grant Information: R01 AG084098 United States AG NIA NIH HHS; R01 GM145960 United States GM NIGMS NIH HHS; ZIA ES103341 United States ImNIH Intramural NIH HHS; ZIC ES103326 United States ImNIH Intramural NIH HHS
Substance Nomenclature: 9013-31-4 (Apoferritins)
0 (Liposomes)
Entry Date(s): Date Created: 20260923 Date Completed: 20260923 Latest Revision: 20260926
Update Code: 20260926
PubMed Central ID: PMC13600247
DOI: 10.1126/sciadv.aeh5900
PMID: 42777058
Database: MEDLINE
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  Data: Acoustofluidic cryo-EM enables in situ particle manipulation for uniform, high-quality cryo-EM specimens.
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  Data: <searchLink fieldCode="AU" term="%22Yang+K%22">Yang K</searchLink>; Thomas Lord Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA.<br /><searchLink fieldCode="AU" term="%22Peele+W%22">Peele W</searchLink>; Genome Integrity and Structural Biology Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Department of Health and Human Services, Research Triangle Park, Durham, NC 27709, USA.<br /><searchLink fieldCode="AU" term="%22Liu+P%22">Liu P</searchLink>; Thomas Lord Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA.<br /><searchLink fieldCode="AU" term="%22Xia+J%22">Xia J</searchLink>; Thomas Lord Department of Mechanical Engineering and Materials Science, Duke University, Durham, NC 27708, USA.<br /><searchLink fieldCode="AU" term="%22Dandey+V%22">Dandey V</searchLink>; Genome Integrity and Structural Biology Laboratory, National Institute of Environmental Health Sciences, National Institutes of Health, Department of Health and Human Services, Research Triangle Park, Durham, NC 27709, USA.<br /><searchLink fieldCode="AU" term="%22Zhong+R%22">Zhong R</searchLink>; 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  Data: Cryo-electron microscopy (cryo-EM) has revolutionized structural biology by enabling imaging of macromolecular complexes at near-atomic resolution, but its success depends critically on specimen quality. Standard plunge freezing often produces nonuniform particle distributions, resulting in poor particle statistics and reduced imaging quality, particularly for vesicular structures and low-concentration, labile macromolecular complexes. Here, we introduce acoustofluidic cryo-EM (ACE), a noncontact specimen-preparation method that integrates acoustofluidic manipulation into a conventional plunge-freezing workflow. ACE enables localized nanoscale particle manipulation by mechanically coupling acoustic excitation into the cryo-EM grid, where forced vibration generates localized acoustic streaming through viscous boundary-layer effects. Guided by this physical mechanism, validation using liposomes and apoferritin demonstrates that ACE increases particle density by up to fourfold while preserving structural integrity and improving imaging quality. This strategy provides a broadly applicable approach for enhancing cryo-EM specimen quality, improving imaging efficiency, and facilitating high-resolution structural analysis.
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  Label: References
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