IP Library › Granted Patent US 12,469,670
Granted Patent B2
US 12,469,670 · App. 17/914,188 · Granted Nov 11, 2025

Electron microscopy support

Inventors: Christopher J. Russo (Cambridge, GB); Katerina Naydenova (Cambridge, GB)
Assignee: United Kingdom Research and Innovation
H01J37/20H01J37/28H01J2237/2001
View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 12,469,670
App. No.
17/914,188
Granted
Nov 11, 2025
Kind
B2
Abstract

A support for an electron microscopy sample, the support comprising a metallic foil having one or more holes therethrough wherein thickness of the metallic foil is less than 50 nm and/or the mean linear intercept grain size is 50 nm or less, wherein the ratio of the diameter of each hole to the thickness of the metallic foil is 15:1 or less, and wherein the metallic foil consists of either (a) one or more metals selected from transition metals, aluminium and beryllium, or an alloy thereof; or (b) degenerately doped silicon wherein the dopant element is selected from boron, aluminium, boron and arsenic at a concentration of 10 20 atoms/cm 3 or higher.

Claims (17)

1 . A support for an electron microscopy sample, the support comprising a metallic foil having one or more holes therethrough wherein thickness of the metallic foil is less than 50 nm and/or the mean linear intercept grain size is 50 nm or less, wherein the ratio of the diameter of each hole to the thickness of the metallic foil is 15:1 or less, and wherein the metallic foil consists of either (a) one or more metals selected from transition metals, aluminium and beryllium, or an alloy thereof; or (b) degenerately doped silicon wherein the dopant element is selected from boron, aluminium, boron and arsenic at a concentration of 10 20 atoms/cm 3 or higher.

2 . A support according to claim 1 wherein thickness of the metallic foil is less than 50 nm and the mean linear intercept grain size is 50 nm or less.

3 . A support according to claim 1 wherein the edge roughness of each hole is 20 nm or less as measured by the root mean square deviation from the expected theoretical hole edge profile.

4 . A support according to claim 1 wherein the diameter of each hole is 750 nm or less.

5 . A support according to claim 1 wherein the support has a light wavelength transmittance maximum of from 650 to 800 nm.

6 . A support according to claim 1 wherein the holes are arranged in a hexagonal array or a square pattern array.

7 . A support according to claim 1 wherein the metallic foil is suspended across holes in an electron microscopy grid.

8 . A support according to claim 7 wherein the metallic foil and the grid are integrally formed.

9 . A support according to claim 7 wherein the grid comprises a mesh having a mean hole size that is on a micrometre scale and the mesh holes are tessellating hexagons or tessellating squares.

10 . A support according to claim 1 wherein the metallic foil consists of one or more of gold, palladium and platinum or an alloy thereof, optionally wherein the metallic foil consists of gold or an alloy thereof.

11 . A support according to claim 1 wherein the support consists of one or more of gold, palladium and platinum or an alloy thereof, optionally wherein the support consists of gold or an alloy thereof.

12 . A method of manufacturing a metallic foil for a support according to claim 1 , the method comprising the steps of depositing a metallic layer onto a patterned substrate that is cooled to 200K or less to form a layer having a thickness of 50 nm or less and having one or more holes therethrough; removing the deposited metallic layer; and forming the metallic layer into a support for an electron microscopy sample, wherein the metallic foil consists of either (a) one or more metals selected from transition metals, aluminium and beryllium, or an alloy thereof; or (b) degenerately doped silicon wherein the dopant element is selected from boron, aluminium, boron and arsenic at a concentration of 10 20 atoms/cm 3 or higher.

13 . A method of electron microscopy imaging comprising a step of sequentially imaging a sample suspended in a hole of a support according to claim 1 , wherein each image encompass at least a part of the edge of the hole and the electron beam encompasses the hole and the complete edge of the hole.

14 . A method of electron microscopy imaging according to claim 13 wherein at least a part of the edge of the hole in each image is compared to the other images to remove any relative shift between sequential images and/or wherein the sequential images of the specimen in the hole are weighted to account for damage to the specimen.

15 . A support according to claim 1 wherein the metallic foil and the support each independently consists of one or more of gold, palladium and platinum or an alloy thereof.

16 . A support according to claim 1 wherein the metallic foil and the support both consist of the same material selected from one or more of gold, palladium and platinum or an alloy thereof.

17 . A support according to claim 1 wherein the metallic foil and the support both consist of gold or an alloy thereof.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jan 26, 2023
From: RUSSO, CHRISTOPHER J.; NAYDENOVA, KATERINA
To: UNITED KINGDOM RESEARCH AND INNOVATION
Reel/Frame 062497/0710 →
Priority Claims (1)
GB 2004272 · Mar 24, 2020 · national
Continuity (1)
Related Publication 20230131360A1 · Apr 27, 2023
References Cited (90)
US 5369241A · Taylor et al. · 1994 [cited by applicant]
US 5369336A · Koinuma et al. · 1994 [cited by applicant]
US 5369337A · Yanagi et al. · 1994 [cited by applicant]
US 5369947A · Dummersdorf et al. · 1994 [cited by applicant]
US 5370767A · Miyakuni et al. · 1994 [cited by applicant]
US 5372199A · Cegielski et al. · 1994 [cited by applicant]
US 5372674A · Steinberg · 1994 [cited by applicant]
US 5372862A · Krishnaswamy · 1994 [cited by applicant]
US 5374327A · Imahashi et al. · 1994 [cited by applicant]
US 5376224A · Zarowin · 1994 [cited by applicant]
US 5376332A · Martens et al. · 1994 [cited by applicant]
US 5376591A · Maeda et al. · 1994 [cited by applicant]
US 5414261A · Ellisman · 1995 [cited by examiner]
US 5540959A · Wang · 1996 [cited by applicant]
US 5986264A · Grunewald · 1999 [cited by examiner]
US 6768110B2 · Alani · 2004 [cited by examiner]
US 9947505B2 · Russo · 2018 [cited by examiner]
US 10157725B2 · Passmore · 2018 [cited by examiner]
US 11067486B2 · Lamers · 2021 [cited by examiner]
US 20030127595A1 · Nakamura · 2003 [cited by examiner]
US 20070131873A1 · Allred, Jr. · 2007 [cited by examiner]
US 20100081260A1 · Chen · 2010 [cited by applicant]
US 20100243482A1 · Chao · 2010 [cited by examiner]
US 20110200787A1 · Regan · 2011 [cited by examiner]
US 20120104253A1 · Tsuneta et al. · 2012 [cited by applicant]
US 20130043387A1 · Own · 2013 [cited by examiner]
US 20140014495A1 · Dickinson et al. · 2014 [cited by applicant]
US 20140054458A1 · Own · 2014 [cited by examiner]
US 20160189919A1 · Passmore · 2016 [cited by examiner]
US 20160203942A1 · Russo et al. · 2016 [cited by applicant]
US 20170271126A1 · Ji · 2017 [cited by examiner]
US 20180017558A1 · Terfort et al. · 2018 [cited by applicant]
US 20180250704A1 · Truica-Marasescu et al. · 2018 [cited by applicant]
US 20200012121A1 · Wang · 2020 [cited by examiner]
US 20220185674A1 · Russo et al. · 2022 [cited by applicant]
US 20230139070A1 · Matousek · 2023 [cited by examiner]
US 20230223231A1 · Bischoff · 2023 [cited by examiner]
CN 105000552A · 2015 [cited by applicant]
CN 105531792A · 2016 [cited by applicant]
CN 108660426A · 2018 [cited by applicant]
JP 2013239265A · 2013 [cited by applicant]
JP 2016532267A · 2016 [cited by applicant]
JP 2016534508A · 2016 [cited by applicant]
WO 2006119521A1 · 2006 [cited by applicant]
WO 2011007492A1 · 2011 [cited by applicant]
WO 2011103424A2 · 2011 [cited by applicant]
WO 2013140822A1 · 2013 [cited by applicant]
WO 2014065665A1 · 2014 [cited by applicant]
WO 2020041202A1 · 2020 [cited by applicant]
WO 2020193719A1 · 2020 [cited by applicant]
Marr, et al., “Fabrication of carbon films with ˜500 nm holes for cryo-EM with a direct detector device”, Journal of Structural Biology 185 (2014) 42-47. [cited by applicant]
Russo, et al., “Ultrastable gold substrates: Properties of a support for high-resolution electron cryomicroscopy of biological specimens”, Journal of Structural Biology 193 (2016) 33-44. [cited by applicant]
Huang, X. et al., “Amorphous nickel titanium alloy film: a new choice for cryo electron microscopy sample preparation, ” bioRxiv, Posted Mar. 2, 2020, doi: https://doi.org/10. 1101/2020.03.02.963959; 28 pages. [cited by applicant]
Jia, P. et al., “Large-area freestanding gold nanomembranes with nanoholes,” Mater. Horiz., vol. 6; 1005-1012 (2019). [cited by applicant]
Russo, C.J. and Passmore, L.A., “Ultrastable gold substrates for electron cryomicroscopy,” Science, vol. 346; Issue 6215; 1377-1380 (2014). [cited by applicant]
International Search Report and Written Opinion for International Application No. PCT/EP2021/057628, mailed Jun. 30, 2021. [cited by applicant]
Search Report for GB Application No. GB2004272.7, issued Mar. 10, 2021. [cited by applicant]
Ermantraut, E. et al., “Perforated support foils with pre-defined hole size, shape and arrangement,” Ultramicroscopy, 74: 75-81 (1998). [cited by applicant]
Grant-Jacob, J.A. et al., “Design and fabrication of a 3D-structured gold film with nanopores for local electric field enhancement in the pore,” Nanotechnology, 27: 1-11 (2018). [cited by applicant]
Janbroers, S. et al., “Preparation of carbon-free TEM microgrids by metal sputtering,” Ultramicroscopy, 109: 1105-1109 (2009). [cited by applicant]
Naydenova, K. et al., “Multifunctional graphene supports for electron cryomicroscopy,” PNAS, vol. 116, No. 24: 11718-11724 (2019). [cited by applicant]
A. Einstein, Über die von der molekularkinetischen Theorie der Wärme geforderte Bewegung von in ruhenden Flüssigkeiten suspendierten Teilchen, Ann. Phys. (1905), 549-560, 322. [cited by applicant]
A. J. Noble, et al., Routine single particle CryoEM sample and grid characterization by tomography, May 29, 2018, eLife, 1-42, 7: e34257. [cited by applicant]
A. K. Geim, Graphene: Status and Prospects, Jun. 19, 2009, Science, 1530-1534, 324. [cited by applicant]
C. A. Schneider, et al., NIH Image to ImageJ: 25 years of Image Analysis, Nat. Methods (Jul. 2012), 671-675, 9(7). [cited by applicant]
C. F. Hryc, et al., Accurate model annotation of a near-atomic resolution cryo-EM map, Mar. 21, 2017, Proc Natl Acad Sci USA, 3103-3108, 114: 12. [cited by applicant]
C. J. Russo, et al., A precision cryostat design for manual and semi-automated cryo-plunge instruments, Review of Scientific Instruments (Nov. 2016), 114302, 87(11). [cited by applicant]
C. J. Russo, L. A. Passmore, Controlling protein adsorption on graphene for cryo-EM using low-energy hydrogen plasmas, Nat Methods. Jun. 2014, 649-652, 11(6). [cited by applicant]
C. J. Russo, L. A. Passmore, Progress towards an optimal specimen support for electron cryomicroscopy, Current Opinion in Structural Biology (2016), 81-89, 37. [cited by applicant]
D. N. Mastronarde, S. R. Held, Automated Tilt Series Alignment and Tomographic Reconstruction in IMOD, Journal of Structural Biology (2017), 102-113, 197(2). [cited by applicant]
Definition of manifold, accessed online at https://www.merriam-webster.com/dictionary/manifold on Nov. 4, 2024 (Year: 2024). [cited by applicant]
E. F. Pettersen, et al., UCSF Chimera—A Visualization System for Exploratory Research and Analysis, J. Comput. Chem. (2004), 1605-1612, 25. [cited by applicant]
Hernandez et al.: “Controlling the local chemical reactivity of graphene through spacial functionalization”, Carbon, Elsevier, Oxford, GB, vol. 60, Apr. 9, 2013 (Apr. 9, 2013), pp. 84-93. [cited by applicant]
J. Dubochet, et al., Cryo-electron microscopy of vitrified specimens, Q. Rev. Biophys. (1988), 129-228, 21, 2. [cited by applicant]
J. Hattne, et al., Analysis of Global and Site-Specific Radiation Damage in Cryo-EM, May 1, 2018, Structure, 759-766, 26. [cited by applicant]
J. Zivanov, et al., A Bayesian approach to beam-induced motion correction in cryo-EM single-particle analysis, IUCrJ (2019), 5-17, 6. [cited by applicant]
J. Zivanov, et al., New tools for automated high-resolution cryo-EM structure determination in RELION-3, Nov. 9, 2018, eLife, 1-22, 7: e42166. [cited by applicant]
K. Naydenova, C. J. Russo, Determining and Controlling the Surface Properties of Cryo-EM Specimen Supports, Microscopy and Microanalysis (2018), 880-881, 24 (Suppl 1). [cited by applicant]
K. Naydenova, C. J. Russo, Measuring the effects of particle orientation to improve the efficiency of electron cryomicroscopy, Sep. 20, 2017, Nature Communications, 1-5, 8:62. [cited by applicant]
Kai Zhang, Gctf: Real-time CTF determination and correction, Journal of Structural Biology (2016), 1-12, 193. [cited by applicant]
Katerina Naydenova et al.: “Patterned Functionalization of Graphene specimen for CryoEM with a Low-energy Plasma Instrument”, Science, Jan. 1, 2014 (Jan. 1, 2014), pp. 1377-1380. [cited by applicant]
Naydenova, et al., “Patterned Functionalization of Graphene Specimen Supports for CryoEM with a Low-Energy Plasma Instrument,” post-deadline poster presentation at the Microscopy & Microanalysis 2019 Conference, Aug. 5,… [cited by applicant]
Naydenova, K. et al., “Multifunctional graphene supports for electron cryomicroscopy,” Proceedings of the National Academy of Sciences, vol. 116; No. 24; 11718-11724; Includes Suppl Material (2019). [cited by applicant]
Pantelic, R. et al., “The application of Graphene as a sample support in Transmission Electron Microscopy,” Support in Transmission Electron Microscopy, Solid State Communications, Elsevier, p. 1375-1382 (2012). [cited by applicant]
Peter B. Rosenthal and Richard Henderson, Optimal Determination of Particle Orientation, Absolute Hand, and Contrast Loss in Single-particle Electron Cryomicroscopy, J. Mol. Biol. (2003), 721-745, 333. [cited by applicant]
R. M. Glaeser, Proteins, interfaces, and cryo-EM grids, Current Opinion in Colloid and Interface Science (2018), 1-11, 34. [cited by applicant]
Search Report for GB Application No. GB1904187.0, Graphene Functionalization Method, Apparatus, and Functionalized Graphene Product dated Aug. 6, 2019. [cited by applicant]
T. Hussain, et al., Large-Scale Movements of IF3 and tRNA during Bacterial Translation Initiation, Cell (2016), 133-144, 167. [cited by applicant]
Table of Contents, Science 2014; 343: 3-4 (Jan. 3, 2014) (Year: 2014). [cited by applicant]
X. Li, et al., Electron counting and beam-induced motion correction enable near atomic resolution single particle cryoEM, Nat. Methods (Jun. 2013), 584-590, 10(6). [cited by applicant]