Abstract
3D correlative microscopy methods have revolutionized biomedical research, allowing the acquisition of multidimensional information to gain an in-depth understanding of biological systems. With the advent of relevant cryo-preservation methods, correlative imaging of cryogenically preserved samples has led to nanometer resolution imaging (2–50 nm) under harsh imaging regimes such as electron and soft X-ray tomography. These methods have now been combined with conventional and super-resolution fluorescence imaging at cryogenic temperatures to augment information content from a given sample, resulting in the immediate requirement for protocols that facilitate hassle-free, unambiguous cross-correlation between microscopes. We present here sample preparation strategies and a direct comparison of different working fiducialization regimes that facilitate 3D correlation of cryo-structured illumination microscopy and cryo-soft X-ray tomography. Our protocol has been tested at two synchrotron beamlines (B24 at Diamond Light Source in the UK and BL09 Mistral at ALBA in Spain) and has led to the development of a decision aid that facilitates experimental design with the strategic use of markers based on project requirements. This protocol takes between 1.5 h and 3.5 d to complete, depending on the cell populations used (adherent cells may require several days to grow on sample carriers).
This is a preview of subscription content, access via your institution
Access options
Access Nature and 54 other Nature Portfolio journals
Get Nature+, our best-value online-access subscription
$29.99 / 30 days
cancel any time
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
Data availability
Original imaging data referenced in the manuscript are deposited at the BioImage Archive (https://www.ebi.ac.uk/biostudies/BioImages) and EMPIAR (https://www.ebi.ac.uk/pdbe/emdb/empiar/). The accession numbers for the data deposited at EMPIAR are EMPIAR-10617, EMPIAR-10618, EMPIAR-10619, EMPIAR-10620, EMPIAR-10621, EMPIAR-10622 and EMPIAR-10624, and the accession numbers for the data deposited at the BioImage Archive are S-BIAD36, S-BIAD37, S-BIAD38, S-BIAD39, S-BIAD40, S-BIAD41 and S-BSST576.
References
Peddie, C. J. & Schieber, N. L. The importance of sample processing for correlative imaging (or, rubbish in, rubbish out). In Correlative Imaging (eds. Verkade, P. & Collinson, L.) 37–66 (John Wiley & Sons, 2020).
Sochacki, K. A., Shtengel, G., van Engelenburg, S. B., Hess, H. F. & Taraska, J. W. Correlative super-resolution fluorescence and metal-replica transmission electron microscopy. Nat. Methods 11, 305–308 (2014).
Löschberger, A., Franke, C., Krohne, G., van de Linde, S. & Sauer, M. Correlative super-resolution fluorescence and electron microscopy of the nuclear pore complex with molecular resolution. J. Cell Sci. 127, 4351–4355 (2014).
Müller-Reichert, T., Srayko, M., Hyman, A., O’Toole, E. T. & McDonald, K. Correlative light and electron microscopy of early Caenorhabditis elegans embryos in mitosis. Methods Cell Biol. 79, 101–119 (2007).
Watari, N. & Herman, L. Correlative light and electron microscopy of bat islets of Langerhans in hibernating and nonhibernating states. Am. Zoolog. 5, 678 (1965).
Timmermans, F. J. & Otto, C. Contributed review: review of integrated correlative light and electron microscopy. Rev. Sci. Instrum. 86, 011501 (2015).
Betzig, E. et al. Imaging intracellular fluorescent proteins at nanometer resolution. Science 313, 1642–1645 (2006).
Jahn, K. A. et al. Correlative microscopy: providing new understanding in the biomedical and plant sciences. Micron 43, 565–582 (2012).
Guérin, C. J., Liv, N. & Klumperman, J. It’s a small, small world. In Correlative Imaging (eds. Verkade, P. & Collinson, L.) 1–21 (John Wiley & Sons, 2020).
Dubochet, J., McDowall, A. W., Menge, B., Schmid, E. N. & Lickfeld, K. G. Electron microscopy of frozen-hydrated bacteria. J. Bacteriol. 155, 381–390 (1983).
Sartori, A. et al. Correlative microscopy: bridging the gap between fluorescence light microscopy and cryo-electron tomography. J. Struct. Biol. 160, 135–145 (2007).
Schwartz, C. L., Sarbash, V. I., Ataullakhanov, F. I., McIntosh, J. R. & Nicastro, D. Cryo-fluorescence microscopy facilitates correlations between light and cryo-electron microscopy and reduces the rate of photobleaching. J. Microsc. 227, 98–109 (2007).
Bharat, T. A. M. & Kukulski, W. Cryo-correlative light and electron microscopy: toward in situ instructional biology. In Correlative Imaging (eds. Verkade, P. & Collinson, L.) 137–153 (John Wiley & Sons, 2020).
Hampton, C. M. et al. Correlated fluorescence microscopy and cryo-electron tomography of virus-infected or transfected mammalian cells. Nat. Protoc. 12, 150–167 (2017).
Henderson, R. et al. Model for the structure of bacteriorhodopsin based on high-resolution electron cryo-microscopy. J. Mol. Biol. 213, 899–929 (1990).
Hoffman, D. P. et al. Correlative three-dimensional super-resolution and block-face electron microscopy of whole vitreously frozen cells. Science 367, eaaz5357 (2020).
Lučić, V., Rigort, A. & Baumeister, W. Cryo-electron tomography: the challenge of doing structural biology in situ. J. Cell Biol. 202, 407–419 (2013).
Beck, M. & Baumeister, W. Cryo-electron tomography: can it reveal the molecular sociology of cells in atomic detail? Trends Cell Biol. 26, 825–837 (2016).
Schneider, G. Cryo X-ray microscopy with high spatial resolution in amplitude and phase contrast. Ultramicroscopy 75, 85–104 (1998).
Schneider, G. et al. Three-dimensional cellular ultrastructure resolved by X-ray microscopy. Nat. Methods 7, 985–987 (2010).
Groen, J., Conesa, J. J., Valcárcel, R. & Pereiro, E. The cellular landscape by cryo soft X-ray tomography. Biophys. Rev. 11, 611–619 (2019).
Kounatidis, I. et al. 3D correlative cryo-structured illumination fluorescence and soft x-ray microscopy elucidates reovirus intracellular release pathway. Cell 182, 1–16 (2020).
Le Gros, M. A. et al. Biological soft X-ray tomography on beamline 2.1 at the Advanced Light Source. J. Synchrotron Radiat. 21, 1370–1377 (2014).
Sorrentino, A. et al. MISTRAL: a transmission soft X-ray microscopy beamline for cryo nano-tomography of biological samples and magnetic domains imaging. J. Synchrotron Radiat. 22, 1112–1117 (2015).
Balint, S. et al. Supramolecular attack particles are autonomous killing entities released from cytotoxic T cells. Science 368, 897–901 (2020).
Phillips, M. et al. CryoSIM: super resolution 3D structured illumination cryogenic fluorescence microscopy for correlated ultra-structural imaging. Optica 7, 802–812 (2020).
Kaufmann, R., Hagen, C. & Grünewald, K. Super-resolution fluorescence microscopy of cryo-immobilized samples. In European Microscopy Congress 2016: Proceedings 1017–1017. https://onlinelibrary.wiley.com/doi/abs/10.1002/9783527808465.EMC2016.6928 (2016).
Kaufmann, R. et al. Super-resolution microscopy using standard fluorescent proteins in intact cells under cryo-conditions. Nano Lett. 14, 4171–4175 (2014).
Kaufmann, R., Hagen, C. & Grünewald, K. Fluorescence cryo-microscopy: current challenges and prospects. Curr. Opin. Chem. Biol. 20, 86–91 (2014).
Duke, E. M. H. et al. Imaging endosomes and autophagosomes in whole mammalian cells using correlative cryo-fluorescence and cryo-soft X-ray microscopy (cryo-CLXM). Ultramicroscopy 143, 77–87 (2014).
Fokkema, J. et al. Fluorescently labelled silica coated gold nanoparticles as fiducial markers for correlative light and electron microscopy. Sci. Rep. 8, 1–10 (2018).
Geissinger, H. D. A precise stage arrangement for correlative microscopy for specimens mounted on glass slides, stubs or EM grids. J. Microsc. 100, 113–117 (1974).
Su, Y. et al. Multi-dimensional correlative imaging of subcellular events: combining the strengths of light and electron microscopy. Biophys. Rev. 2, 121–135 (2010).
Lakowicz, J. R. Fluorophores. In Principles of Fluorescence Spectroscopy. 3rd edn, 63–95 (Springer, 2006).
Lavis, L. D. & Raines, R. T. Bright ideas for chemical biology. ACS Chem. Biol. 3, 142–155 (2008).
Anderson, K., Nilsson, T. & Fernandez-Rodriguez, J. Challenges for CLEM from a light microscopy perspective. In Correlative Imaging (eds. Verkade, P. & Collinson, L.) 23–35 (John Wiley & Sons, 2020).
Paul-Gilloteaux, P. & Schorb, M. Correlating data from imaging modalities. In Correlative Imaging (eds. Verkade, P. & Collinson, L.) 191–210 (John Wiley & Sons, 2020).
Pereiro, E., Chichón, F. J. & Carrascosa, J. L. Correlative cryo soft X-ray imaging. In Correlative Imaging (eds. Verkade, P. & Collinson, L.) 155–169 (John Wiley & Sons, 2020).
Rizk, A. et al. Segmentation and quantification of subcellular structures in fluorescence microscopy images using Squassh. Nat. Protoc. 9, 586–596 (2014).
Pereiro, E., Nicolás, J., Ferrer, S. & Howells, M. R. A soft X-ray beamline for transmission X-ray microscopy at ALBA. J. Synchrotron Radiat. 16, 505–512 (2009).
Harkiolaki, M. et al. Cryo-soft X-ray tomography: using soft X-rays to explore the ultrastructure of whole cells. Emerg. Top. Life Sci. 2, 81–92 (2018).
Gustafsson, M. G. L. et al. Three-dimensional resolution doubling in wide-field fluorescence microscopy by structured illumination. Biophys. J. 94, 4957–4970 (2008).
Carrascosa, J. L. et al. Cryo-X-ray tomography of vaccinia virus membranes and inner compartments. J. Struct. Biol. 168, 234–239 (2009).
Pérez-Berná, A. J. et al. Structural changes in cells imaged by soft X-ray cryo-tomography during hepatitis C virus infection. ACS Nano 10, 6597–6611 (2016).
Spink, M. C. et al. Correlation of cryo soft X-ray tomography with cryo fluorescence microscopy to characterise cellular organelles at beamline B24, Diamond Light Source. Microsc. Microanal. 24, 374–375 (2018).
Bohren, C. F. & Huffman, D. R. Absorption and Scattering of Light by Small Particles (John Wiley & Sons, 2008).
Kerker, M. The Scattering of Light and Other Electromagnetic Radiation (Academic Press, 2013).
Kreibig, U. & Vollmer, M. Optical Properties of Metal Clusters (Springer Science & Business Media, 2013).
Papavassiliou, G. C. Optical properties of small inorganic and organic metal particles. Prog. Solid State Chem. 12, 185–271 (1979).
Weiner, A. et al. Vitrification of thick samples for soft X-ray cryo-tomography by high pressure freezing. J. Struct. Biol. 181, 77–81 (2013).
Gal, A. et al. Native-state imaging of calcifying and noncalcifying microalgae reveals similarities in their calcium storage organelles. Proc. Natl Acad. Sci. USA 115, 11000–11005 (2018).
Conesa, J. J. et al. Unambiguous intracellular localization and quantification of a potent iridium anticancer compound by correlative 3D cryo X-ray imaging. Angew. Chem. Int. Ed. Engl. 59, 1270–1278 (2020).
Ando, T. et al. The 2018 correlative microscopy techniques roadmap. J. Phys. D Appl. Phys. 51, 443001 (2018).
Arnold, J. et al. Site-specific cryo-focused ion beam sample preparation guided by 3D correlative microscopy. Biophys. J. 110, 860–869 (2016).
Kukulski, W. et al. Correlated fluorescence and 3D electron microscopy with high sensitivity and spatial precision. J. Cell Biol. 192, 111–119 (2011).
de Boer, P., Hoogenboom, J. P. & Giepmans, B. N. G. Correlated light and electron microscopy: ultrastructure lights up! Nat. Methods 12, 503–513 (2015).
Varsano, N. et al. Development of correlative cryo-soft X-ray tomography and stochastic reconstruction microscopy. A study of cholesterol crystal early formation in cells. J. Am. Chem. Soc. 138, 14931–14940 (2016).
Hagen, C. et al. Multimodal nanoparticles as alignment and correlation markers in fluorescence/soft X-ray cryo-microscopy/tomography of nucleoplasmic reticulum and apoptosis in mammalian cells. Ultramicroscopy 146, 46–54 (2014).
Elgass, K. D., Smith, E. A., LeGros, M. A., Larabell, C. A. & Ryan, M. T. Analysis of ER-mitochondria contacts using correlative fluorescence microscopy and soft X-ray tomography of mammalian cells. J. Cell Sci. 128, 2795–2804 (2015).
McDermott, G., Le Gros, M. A., Knoechel, C. G., Uchida, M. & Larabell, C. A. Soft X-ray tomography and cryogenic light microscopy: the cool combination in cellular imaging. Trends Cell Biol. 19, 587–595 (2009).
Kapishnikov, S. et al. Unraveling heme detoxification in the malaria parasite by in situ correlative X-ray fluorescence microscopy and soft X-ray tomography. Sci. Rep. 7, 7610 (2017).
Smith, E. A. et al. Quantitatively imaging chromosomes by correlated cryo-fluorescence and soft x-ray tomographies. Biophys. J. 107, 1988–1996 (2014).
van Hest, J. J. Ha et al. Towards robust and versatile single nanoparticle fiducial markers for correlative light and electron microscopy. J. Microsc. 274, 13–22 (2019).
Aslan, K., Wu, M., Lakowicz, J. R. & Geddes, C. D. Fluorescent core-shell Ag@SiO2 nanocomposites for metal-enhanced fluorescence and single nanoparticle sensing platforms. J. Am. Chem. Soc. 129, 1524–1525 (2007).
Aslan, K. & Geddes, C. D. Metal-enhanced fluorescence: progress towards a unified plasmon-fluorophore description. In Metal-Enhanced Fluorescence (ed. Geddes, C. D.) 1–23 (John Wiley & Sons, 2010).
Lee, D., Lee, J., Song, J., Jen, M. & Pang, Y. Homogeneous silver colloidal substrates optimal for metal-enhanced fluorescence. Phys. Chem. Chem. Phys. 21, 11599–11607 (2019).
Hodgson, L., Verkade, P. & Yamauchi, Y. Correlative light and electron microscopy of influenza virus entry and budding. Influenza Virus (ed. Yamauchi, Y.) 237–260 (Humana Press, 2018).
McGorty, R., Kamiyama, D. & Huang, B. Active microscope stabilization in three dimensions using image correlation. Opt. Nanoscopy https://doi.org/10.1186/2192-2853-2-3 (2013).
Metskas, L. A. & Briggs, J. A. G. Fluorescence-based detection of membrane fusion state on a cryo-EM grid using correlated cryo-fluorescence and cryo-electron microscopy. Microsc. Microanal. 25, 942–949 (2019).
Walling, M. A., Novak, J. A. & Shepard, J. R. E. Quantum dots for live cell and in vivo imaging. Int. J. Mol. Sci. 10, 441–491 (2009).
Wegner, K. D. & Hildebrandt, N. Quantum dots: bright and versatile in vitro and in vivo fluorescence imaging biosensors. Chem. Soc. Rev. 44, 4792–4834 (2015).
Hemelaar, S. R. et al. Nanodiamonds as multi-purpose labels for microscopy. Sci. Rep. 7, 1–9 (2017).
Schade, A. E. et al. Dasatinib, a small-molecule protein tyrosine kinase inhibitor, inhibits T-cell activation and proliferation. Blood 111, 1366–1377 (2008).
Trickett, A. & Kwan, Y. L. T cell stimulation and expansion using anti-CD3/CD28 beads. J. Immunol. Methods 275, 251–255 (2003).
Paul-Gilloteaux, P. et al. eC-CLEM: flexible multidimensional registration software for correlative microscopies. Nat. Methods 14, 102–103 (2017).
Bogovic, J. A., Hanslovsky, P., Wong, A. & Saalfeld, S. Robust registration of calcium images by learned contrast synthesis. In 2016 IEEE 13th International Symposium on Biomedical Imaging (ISBI) 1123–1126 (IEEE, 2016).
Miles, B. T. et al. Direct evidence of lack of colocalisation of fluorescently labelled gold labels used in correlative light electron microscopy. Sci. Rep. 7, 44666 (2017).
Oorschot, V., de Wit, H., Annaert, W. G. & Klumperman, J. A novel flat-embedding method to prepare ultrathin cryosections from cultured cells in their in situ orientation. J. Histochem. Cytochem. 50, 1067–1080 (2002).
Tuijtel, M. W., Koster, A. J., Jakobs, S., Faas, F. G. A. & Sharp, T. H. Correlative cryo super-resolution light and electron microscopy on mammalian cells using fluorescent proteins. Sci. Rep. 9, 1369 (2019).
Schellenberger, P. et al. High-precision correlative fluorescence and electron cryo microscopy using two independent alignment markers. Ultramicroscopy 143, 41–51 (2014).
Pezzi, H. M., Niles, D. J., Schehr, J. L., Beebe, D. J. & Lang, J. M. Integration of magnetic bead-based cell selection into complex isolations. ACS Omega 3, 3908–3917 (2018).
Uludag, H., Ubeda, A. & Ansari, A. At the intersection of biomaterials and gene therapy: progress in non-viral delivery of nucleic acids. Front. Bioeng. Biotechnol. 7, 131 (2019).
Booth, D. G., Beckett, A. J., Prior, I. A. & Meijer, D. SuperCLEM: an accessible correlative light and electron microscopy approach for investigation of neurons and glia in vitro. Biol. Open 8, bio042085 (2019).
Telling, N. D. et al. Iron biochemistry is correlated with amyloid plaque morphology in an established mouse model of Alzheimer’s disease. Cell Chem. Biol. 24, 1205–1215.e3 (2017).
Jamme, F. et al. Synchrotron multimodal imaging in a whole cell reveals lipid droplet core organization. J. Synchrotron Radiat. 27, 772–778 (2020).
Ahn, S., Jung, S. Y. & Lee, S. J. Gold nanoparticle contrast agents in advanced X-ray imaging technologies. Molecules 18, 5858–5890 (2013).
Niclis, J. C. et al. Three-dimensional imaging of human stem cells using soft X-ray tomography. J. R. Soc. Interface 12, 20150252 (2015).
Conesa, J. J. et al. Intracellular nanoparticles mass quantification by near-edge absorption soft X-ray nanotomography. Sci. Rep. 6, 22354 (2016).
Matsuda, A., Schermelleh, L., Hirano, Y., Haraguchi, T. & Hiraoka, Y. Accurate and fiducial-marker-free correction for three-dimensional chromatic shift in biological fluorescence microscopy. Sci. Rep. 8, 7583 (2018).
Schindelin, J. et al. Fiji: an open-source platform for biological-image analysis. Nat. Methods 9, 676–682 (2012).
Ball, G. et al. SIMcheck: a toolbox for successful super-resolution structured illumination microscopy. Sci. Rep. 5, 15915 (2015).
Kremer, J. R., Mastronarde, D. N. & McIntosh, J. R. Computer visualization of three-dimensional image data using IMOD. J. Struct. Biol. 116, 71–76 (1996).
de Chaumont, F. et al. Icy: an open bioimage informatics platform for extended reproducible research. Nat. Methods 9, 690–696 (2012).
Luengo, I. et al. SuRVoS: Super-Region Volume Segmentation workbench. J. Struct. Biol. 198, 43–53 (2017).
Goddard, T. D. et al. UCSF ChimeraX: meeting modern challenges in visualization and analysis. Protein Sci. 27, 14–25 (2018).
Bouterfa, H. et al. Expression of different extracellular matrix components in human brain tumor and melanoma cells in respect to variant culture conditions. J. Neurooncol. 44, 23–33 (1999).
Vaz, F. et al. Accessibility to peptidoglycan is important for the recognition of gram-positive bacteria in Drosophila. Cell Rep. 27, 2480–2492.e6 (2019).
Vizcardo, R. et al. Regeneration of human tumor antigen-specific T cells from iPSCs derived from mature CD8+ T cells. Cell Stem Cell 12, 31–36 (2013).
Peng, T. et al. Determining the distribution of probes between different subcellular locations through automated unmixing of subcellular patterns. Proc. Natl Acad. Sci. USA 107, 2944–2949 (2010).
Farmer, B. C., Kluemper, J. & Johnson, L. A. Apolipoprotein E4 alters astrocyte fatty acid metabolism and lipid droplet formation. Cells 8, 182 (2019).
Chazotte, B. Labeling lysosomes in live cells with LysoTracker. Cold Spring Harb. Protoc. 2011, pdb.prot5571 (2011).
Bianchini, P. et al. Live imaging of mammalian retina: rod outer segments are stained by conventional mitochondrial dyes. J. Biomed. Opt. 13, 054017 (2008).
Awasthi, S., Madhusoodhanan, R. & Wolf, R. Surfactant protein-A and toll-like receptor-4 modulate immune functions of preterm baboon lung dendritic cell precursor cells. Cell. Immunol. 268, 87–96 (2011).
Drulyte, I. et al. Approaches to altering particle distributions in cryo-electron microscopy sample preparation. Acta Crystallogr. D Struct. Biol. 74, 560–571 (2018).
Thompson, R. F., Walker, M., Siebert, C. A., Muench, S. P. & Ranson, N. A. An introduction to sample preparation and imaging by cryo-electron microscopy for structural biology. Methods 100, 3–15 (2016).
Grassucci, R. A., Taylor, D. J. & Frank, J. Preparation of macromolecular complexes for cryo-electron microscopy. Nat. Protoc. 2, 3239–3246 (2007).
Hiroyasu, A., DeWitt, D. C. & Goodman, A. G. Extraction of hemocytes from Drosophila melanogaster larvae for microbial infection and analysis. J. Vis. Exp. 135, 57077 (2018).
Dobro, M. J., Melanson, L. A., Jensen, G. J. & McDowall, A. W. Plunge freezing for electron cryomicroscopy. In Cryo-EM, Part A: Sample Preparation and Data Collection Vol. 481 (ed. Jensen, G. J.) 63–82 (Academic Press, 2010).
Noble, A. J. et al. Routine single particle CryoEM sample and grid characterization by tomography. eLife 7, e34257 (2018).
Acknowledgements
We thank P. Paul-Gilloteaux for her invaluable help with eC-CLEM and previous and current members of the B24 and BL09 teams, with special thanks to M. Spink for instrumentation support and A. Taylor and A. Prescott for technical support. We also thank A. Clayton for suggestions in trying new reagents and M. Dumoux for help with laboratory techniques, advice and training. We acknowledge the support of Micron in the development, application and maintenance of the B24 super-resolution facility. We also thank A. Aires Trapote (CIC BiomaGUNE, San Sebastian, Spain), A. V. Villar, A. R. Palanca, D. Maestro Lavín (IBBTEC, Santander, Spain) and J. Conesa (ALBA and CNB-CSIC). This work was carried out with the support of the Diamond Light Source, instrument B24 (proposals MX18737, MX20321, BI22274, BI23046 and BI25162). We acknowledge ALBA for allocated MISTRAL beamtimes 2018093099 and 2019093739. This project has received funding from the European Commission Horizon 2020 iNEXT-Discovery project and the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie grant agreement 75439 and Wellcome awards 091911/Z/11/Z and 107457/Z/15/Z. S.B. is supported by ERC AdG670930.
Author information
Authors and Affiliations
Contributions
C.A.O. coordinated and produced the manuscript with the help of I.K., J.G., E.P. and M.H. C.A.O., I.K., J.G., A.L.C., K.L.N. and S.B. provided data, protocols and critical evaluation of results. M.A.K. and T.M.F. provided support with software and protocol development. I.M.D. supported cryoSIM operations and optimization. E.P. and M.H. managed beamline resources, supervised experiments and evaluated applicability and user-friendliness.
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing interests.
Additional information
Peer review information Nature Protocols thanks Gerd Schneider and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.
Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Related links
Key references using this protocol
Kounatidis, I. et al. Cell 182, 1–16 (2020): https://doi.org/10.1016/j.cell.2020.05.051
Phillips, M. et al. Optica 7, 802–812 (2020): https://doi.org/10.1364/OPTICA.393203
Harkiolaki, M. et al. Emerg. Top. Life Sci. 2, 81–92 (2018): https://doi.org/10.1042/ETLS20170086
Supplementary information
Supplementary Information
Supplementary Note 1, Supplementary Figs. 1–5 and Supplementary Tables 1 and 2.
Rights and permissions
About this article
Cite this article
Okolo, C.A., Kounatidis, I., Groen, J. et al. Sample preparation strategies for efficient correlation of 3D SIM and soft X-ray tomography data at cryogenic temperatures. Nat Protoc 16, 2851–2885 (2021). https://doi.org/10.1038/s41596-021-00522-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/s41596-021-00522-4
This article is cited by
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.