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Journal articles on the topic 'Cryo-CLEM'

Author: Grafiati

Published: 14 September 2024

Last updated: 31 July 2025

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1

Metskas, Lauren Ann, and John A. G. Briggs. "Fluorescence-Based Detection of Membrane Fusion State on a Cryo-EM Grid using Correlated Cryo-Fluorescence and Cryo-Electron Microscopy." Microscopy and Microanalysis 25, no. 4 (2019): 942–49. http://dx.doi.org/10.1017/s1431927619000606.

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AbstractCorrelated light and electron microscopy (CLEM) has become a popular technique for combining the protein-specific labeling of fluorescence with electron microscopy, both at room and cryogenic temperatures. Fluorescence applications at cryo-temperatures have typically been limited to localization of tagged protein oligomers due to known issues of extended triplet state duration, spectral shifts, and reduced photon capture through cryo-CLEM objectives. Here, we consider fluorophore characteristics and behaviors that could enable more extended applications. We describe how dialkylcarbocan

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2

Moser, Felipe, Vojtěch Pražák, Valerie Mordhorst, et al. "Cryo-SOFI enabling low-dose super-resolution correlative light and electron cryo-microscopy." Proceedings of the National Academy of Sciences 116, no. 11 (2019): 4804–9. http://dx.doi.org/10.1073/pnas.1810690116.

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Correlative light and electron cryo-microscopy (cryo-CLEM) combines information from the specific labeling of fluorescence cryo-microscopy (cryo-FM) with the high resolution in environmental context of electron cryo-microscopy (cryo-EM). Exploiting super-resolution methods for cryo-FM is advantageous, as it enables the identification of rare events within the environmental background of cryo-EM at a sensitivity and resolution beyond that of conventional methods. However, due to the need for relatively high laser intensities, current super-resolution cryo-CLEM methods require cryo-protectants o

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3

Li, Shuoguo, Gang Ji, Xiaojun Huang, et al. "A New Solution of Non-integrated Correlative Light and Electron Microscopy Based on High-vacuum Optical Platform." Microscopy and Microanalysis 22, S3 (2016): 248–49. http://dx.doi.org/10.1017/s1431927616002099.

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Abstract Correlative light and electron microscopy (CLEM) offers a means of guiding the search for the unique or rare events by fluorescence microscopy (FM) and allows electron microscopy (EM) to zoom in on them for subsequent EM examination in three-dimensions (3D) and with nanometer-scale resolution. FM visualizes the localization of specific antigens by using fluorescent tags or proteins in a large field-of-view to study their cellular function, whereas EM provides the high level of resolution for complex structures. And cryo CLEM combines the advantages of maintaining structural preservati

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4

Thomas, Connon I., Nicolai T. Urban, Ye Sun, et al. "Cryo-Confocal Imaging for CLEM Mapping in Brain Tissues." Microscopy Today 29, no. 5 (2021): 34–39. http://dx.doi.org/10.1017/s1551929521001073.

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Abstract:In correlative light and electron microscopy (CLEM) workflows, identifying the same sub-cellular features in tissue by both light (LM) and electron microscopy (EM) remains a challenge. Furthermore, use of cryo-fixation for EM is desirable to capture rapid biological phenomena. Here, we describe a workflow that incorporates cryo-confocal laser scanning microscopy into the CLEM process, mapping cells in brain slices to re-image them with serial section scanning electron microscopy (ssSEM) array tomography. The addition of Airyscan detection increased the signal-to-noise ratio (SNR), all

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5

Hampton, Cheri M. "Practical Strategies for cryo-CLEM Experiments." Microscopy and Microanalysis 23, S1 (2017): 1400–1401. http://dx.doi.org/10.1017/s1431927617007668.

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6

Shahmoradian, Sarah. "Elucidating Tau Fibril Formation using Correlative Cryo-CLEM in situ." Structural Dynamics 12, no. 2_Supplement (2025): A351. https://doi.org/10.1063/4.0000657.

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Accurate modeling of tau protein aggregation in diverse cell systems, such as fluorescently tagged tau-expressing cell lines or 'biosensor' cells, is crucial for advancing tauopathy research. This encompasses understanding the early stages of tau fibril formation and localization within these cells, a vital but underexplored aspect in studying these neurodegenerative diseases. Our aim was to elucidate the structure and formation of tau fibrils in fluorescently tagged tau-expressing cell lines and iPSC- derived human neurons, assessing their resemblance to amyloid structures in tauopathies and

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7

Kamp, Arnold, Martijn van Nugteren, Hildo Vader, et al. "Automated Cryo-plunging Robot to Prepare Samples for Single Particle Analysis (SPA), Cryo-EM, Cryo-ET, Cryo-fluorescence and Cryo-CLEM." Microscopy and Microanalysis 26, S2 (2020): 2732–33. http://dx.doi.org/10.1017/s1431927620022606.

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8

Paraan, Reza, Victoria Hewitt, Yusuke Hirabayashi, Franck Polleux, Clint Potter, and Bridget Carragher. "Characterization of ER-mitochondria contact sites using cryo-CLEM." Microscopy and Microanalysis 27, S1 (2021): 1712–13. http://dx.doi.org/10.1017/s1431927621006255.

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9

Schwertner, Michael, and Duncan Stacey. "Cryo-Correlative Light and Electron Microscopy (Cryo-CLEM): Specimen Workflow Paths and Recent Instrument Developments." Microscopy and Microanalysis 21, S3 (2015): 1565–66. http://dx.doi.org/10.1017/s1431927615008600.

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10

Sexton, Danielle L., Steffen Burgold, Andreas Schertel, and Elitza I. Tocheva. "Super-resolution confocal cryo-CLEM with cryo-FIB milling for in situ imaging of Deinococcus radiodurans." Current Research in Structural Biology 4 (2022): 1–9. http://dx.doi.org/10.1016/j.crstbi.2021.12.001.

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11

Shahmoradian, Sarah, Jim Monistrol, Hung Tri Tran, et al. "Abstract 2445 Elucidating Tau Fibril Formation using Correlative Cryo-CLEM in situ." Journal of Biological Chemistry 300, no. 3 (2024): 107098. http://dx.doi.org/10.1016/j.jbc.2024.107098.

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12

de Beer, Marit, Rona Roverts, Xavier Heiligenstein, Edwin Lamers, Nico Sommerdijk, and Anat Akiva. "Visualizing Biological Tissues: A Multiscale Workflow from Live Imaging to 3D Cryo-CLEM." Microscopy and Microanalysis 27, S2 (2021): 11–12. http://dx.doi.org/10.1017/s1431927621013635.

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13

Casanova, G., F. Nolin, L. Wortham, D. Ploton, V. Banchet, and J. Michel. "Shrinkage of freeze-dried cryosections of cells: Investigations by EFTEM and cryo-CLEM." Micron 88 (September 2016): 77–83. http://dx.doi.org/10.1016/j.micron.2016.06.005.

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14

Capitanio, Cristina, Anna Bieber, and Florian Wilfling. "How Membrane Contact Sites Shape the Phagophore." Contact 6 (January 2023): 251525642311624. http://dx.doi.org/10.1177/25152564231162495.

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During macroautophagy, phagophores establish multiple membrane contact sites (MCSs) with other organelles that are pivotal for proper phagophore assembly and growth. In S. cerevisiae, phagophore contacts have been observed with the vacuole, the ER, and lipid droplets. In situ imaging studies have greatly advanced our understanding of the structure and function of these sites. Here, we discuss how in situ structural methods like cryo-CLEM can give unprecedented insights into MCSs, and how they help to elucidate the structural arrangements of MCSs within cells. We further summarize the current k

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15

Kim, Hong-Lim, Tae-Ryong Riew, Jieun Park, Youngchun Lee, and In-Beom Kim. "Correlative Light and Electron Microscopy Using Frozen Section Obtained Using Cryo-Ultramicrotomy." International Journal of Molecular Sciences 22, no. 8 (2021): 4273. http://dx.doi.org/10.3390/ijms22084273.

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Immuno-electron microscopy (Immuno-EM) is a powerful tool for identifying molecular targets with ultrastructural details in biological specimens. However, technical barriers, such as the loss of ultrastructural integrity, the decrease in antigenicity, or artifacts in the handling process, hinder the widespread use of the technique by biomedical researchers. We developed a method to overcome such challenges by combining light and electron microscopy with immunolabeling based on Tokuyasu’s method. Using cryo-sectioned biological specimens, target proteins with excellent antigenicity were first i

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16

Bieber, Anna, Cristina Capitanio, Oda Schiøtz, et al. "Precise 3D-correlative FIB-milling of biological samples using METEOR, an integrated cryo-CLEM imaging system." Microscopy and Microanalysis 27, S1 (2021): 3230–32. http://dx.doi.org/10.1017/s1431927621011132.

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17

Jonker, Caspar, Daan Boltje, Jacob Hoogenboom, et al. "Fluorescence-guided lamella fabrication with ENZEL, an integrated cryogenic CLEM solution for the cryo-electron tomography workflow." Microscopy and Microanalysis 27, S1 (2021): 3234–35. http://dx.doi.org/10.1017/s1431927621011144.

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18

Kapishnikov, Sergey, Kenneth Fahy, William Fyans, Fergal O’Reilly, Tony McEnroe, and Paul Sheridan. "Integration of Laboratory Cryo Soft X-ray Tomography into CLEM Workflows for Multimodal Multiscale Imaging of Bulk Samples." Microscopy and Microanalysis 29, Supplement_1 (2023): 1164. http://dx.doi.org/10.1093/micmic/ozad067.595.

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19

Li, Shuoguo, Gang Ji, Yang Shi, et al. "High-vacuum optical platform for cryo-CLEM (HOPE): A new solution for non-integrated multiscale correlative light and electron microscopy." Journal of Structural Biology 201, no. 1 (2018): 63–75. http://dx.doi.org/10.1016/j.jsb.2017.11.002.

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20

Li, Shuoguo, Xing Jia, Tongxin Niu, et al. "HOPE-SIM, a cryo-structured illumination fluorescence microscopy system for accurately targeted cryo-electron tomography." Communications Biology 6, no. 1 (2023). http://dx.doi.org/10.1038/s42003-023-04850-x.

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AbstractCryo-focused ion beam (cryo-FIB) milling technology has been developed for the fabrication of cryo-lamella of frozen native specimens for study by in situ cryo-electron tomography (cryo-ET). However, the precision of the target of interest is still one of the major bottlenecks limiting application. Here, we have developed a cryo-correlative light and electron microscopy (cryo-CLEM) system named HOPE-SIM by incorporating a 3D structured illumination fluorescence microscopy (SIM) system and an upgraded high-vacuum stage to achieve efficiently targeted cryo-FIB. With the 3D super resoluti

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21

Klumpe, Sven, Herman KH Fung, Sara K. Goetz, et al. "A modular platform for automated cryo-FIB workflows." eLife 10 (December 24, 2021). http://dx.doi.org/10.7554/elife.70506.

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Lamella micromachining by focused ion beam milling at cryogenic temperature (cryo-FIB) has matured into a preparation method widely used for cellular cryo-electron tomography. Due to the limited ablation rates of low Ga+ ion beam currents required to maintain the structural integrity of vitreous specimens, common preparation protocols are time-consuming and labor intensive. The improved stability of new-generation cryo-FIB instruments now enables automated operations. Here, we present an open-source software tool, SerialFIB, for creating automated and customizable cryo-FIB preparation protocol

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22

DeRosier, David J. "Where in the cell is my protein?" Quarterly Reviews of Biophysics 54 (2021). http://dx.doi.org/10.1017/s003358352100007x.

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Abstract The application of cryo-correlative light and cryo-electron microscopy (cryo-CLEM) gives us a way to locate structures of interest in the electron microscope. In brief, the structures of interest are fluorescently tagged, and images from the cryo-fluorescent microscope (cryo-FM) maps are superimposed on those from the cryo-electron microscope (cryo-EM). By enhancing cryo-FM to include single-molecule localization microscopy (SMLM), we can achieve much better localization. The introduction of cryo-SMLM increased the yield of photons from fluorophores, which can benefit localization eff

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23

Fuest, Marie, Miroslava Schaffer, Giovanni Marco Nocera, et al. "In situ Microfluidic Cryofixation for Cryo Focused Ion Beam Milling and Cryo Electron Tomography." Scientific Reports 9, no. 1 (2019). http://dx.doi.org/10.1038/s41598-019-55413-2.

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AbstractWe present a microfluidic platform for studying structure-function relationships at the cellular level by connecting video rate live cell imaging with in situ microfluidic cryofixation and cryo-electron tomography of near natively preserved, unstained specimens. Correlative light and electron microscopy (CLEM) has been limited by the time required to transfer live cells from the light microscope to dedicated cryofixation instruments, such as a plunge freezer or high-pressure freezer. We recently demonstrated a microfluidic based approach that enables sample cryofixation directly in the

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24

Tillu, V. A., G. M. I. Redpath, J. Rae, et al. "Precision in situ cryo-correlative light and electron microscopy of optogenetically-positioned organelles." Journal of Cell Science, September 23, 2024. http://dx.doi.org/10.1242/jcs.262163.

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Unambiguous targeting of cellular structures for in situ cryo-electron microscopy in the heterogeneous, dense, and compacted environment of the cytoplasm remains challenging. Here we have developed a cryogenic correlative light and electron microscopy (cryo-CLEM) workflow which combines thin cells grown on a mechanically defined substratum to rapidly analyse organelles and macromolecular complexes by cryo-electron tomography (cryo-ET). We coupled these advancements with optogenetics to redistribute perinuclear-localised organelles to the cell periphery, allowing visualisation of organelles oth

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25

Kobylynska, M., P. C. Hawes, and R. A. Fleck. "Multi-scale Correlative Workflows, Challenges and Opportunities for Cryo CLEM." Microscopy and Microanalysis 30, Supplement_1 (2024). http://dx.doi.org/10.1093/mam/ozae044.1041.

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26

Klein, Steffen, Benedikt H. Wimmer, Sophie L. Winter, Androniki Kolovou, Vibor Laketa, and Petr Chlanda. "Post-correlation on-lamella cryo-CLEM reveals the membrane architecture of lamellar bodies." Communications Biology 4, no. 1 (2021). http://dx.doi.org/10.1038/s42003-020-01567-z.

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AbstractLamellar bodies (LBs) are surfactant-rich organelles in alveolar cells. LBs disassemble into a lipid-protein network that reduces surface tension and facilitates gas exchange in the alveolar cavity. Current knowledge of LB architecture is predominantly based on electron microscopy studies using disruptive sample preparation methods. We established and validated a post-correlation on-lamella cryo-correlative light and electron microscopy approach for cryo-FIB milled cells to structurally characterize and validate the identity of LBs in their unperturbed state. Using deconvolution and 3D

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27

Gorelick, Sergey, Genevieve Buckley, Gediminas Gervinskas, et al. "PIE-scope, integrated cryo-correlative light and FIB/SEM microscopy." eLife, June 28, 2019. https://doi.org/10.5281/zenodo.3260173.

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Cryo-Electron Tomography (cryo-ET) is emerging as a revolutionary method for resolving the structure of macromolecular complexes <em>in situ.</em> However, sample preparation for <em>in situ </em>Cryo-ET is labour-intensive and can require both cryo-lamella preparation through cryo-Focused Ion Beam (FIB) milling and correlative light microscopy to ensure that the event of interest is present in the lamella. Here, we present an integrated cryo-FIB and light microscope setup called the Photon Ion Electron microscope (PIE-scope) that enables direct and rapid isolation of cellular regions containi

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28

Seewald, Anne, Jingxiao Zhong, Macarena Siri, Peter Fratzl, and Emeline Raguin. "Three-Dimensional Imaging of Vasculature and Forming Quail Femur using Cryo-Correlative Light and Electron Microscopy (cryo-CLEM)." Faraday Discussions, 2025. https://doi.org/10.1039/d5fd00022j.

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Bone mineralization during embryonic development requires the transport and deposition of an enormous amount of mineral precursors. In avian embryos, blood vessels play a dual role in this context: facilitating...

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29

Yang, Suyeon, Machi Takeuchi, Rick R. M. Joosten, John P. M. van Duynhoven, Heiner Friedrich, and Johannes Hohlbein. "Adapting cryogenic correlative light and electron microscopy (cryo-CLEM) for food oxidation studies." Food Structure, February 2024, 100365. http://dx.doi.org/10.1016/j.foostr.2024.100365.

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30

Szabo, Gréta V., and Thomas P. Burg. "Time Resolved Cryo‐Correlative Light and Electron Microscopy." Advanced Functional Materials, April 16, 2024. http://dx.doi.org/10.1002/adfm.202313705.

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AbstractComplex materials exhibit fascinating features especially in situations far from equilibrium. Thus, methods for investigating structural dynamics with sub‐second time resolution are becoming a question of interest at varying spatial scales. With novel microscopy techniques steadily improving, the temporal and spatial limits of multiple imaging methods are investigated with an emphasis on the important role of correlative imaging and cryo‐fixation. A deep‐dive is taken into cryo‐correlative light and electron microscopy (CLEM) as a starting point for multimodal investigations of ultrast

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31

Fu, Xiaofeng, Jiying Ning, Zhou Zhong, Zandrea Ambrose, Simon Charles Watkins, and Peijun Zhang. "AutoCLEM: An Automated Workflow for Correlative Live-Cell Fluorescence Microscopy and Cryo-Electron Tomography." Scientific Reports 9, no. 1 (2019). http://dx.doi.org/10.1038/s41598-019-55766-8.

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AbstractCorrelative light and electron microscopy (CLEM) combines the strengths of both light and electron imaging modalities and enables linking of biological spatiotemporal information from live-cell fluorescence light microscopy (fLM) to high-resolution cellular ultra-structures from cryo-electron microscopy and tomography (cryoEM/ET). This has been previously achieved by using fLM signals to localize the regions of interest under cryogenic conditions. The correlation process, however, is often tedious and time-consuming with low throughput and limited accuracy, because multiple correlation

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32

Bieber, Anna, Cristina Capitanio, Oda Schiøtz, et al. "Precise 3D-correlative FIB-milling of biological samples using METEOR, an integrated cryo-CLEM imaging system." July 30, 2021. https://doi.org/10.1017/s1431927621011132.

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33

Santarella-Mellwig, Rachel, Uta Haselmann, Nicole L. Schieber, et al. "Correlative Light Electron Microscopy (CLEM) for Tracking and Imaging Viral Protein Associated Structures in Cryo-immobilized Cells." Journal of Visualized Experiments, no. 139 (September 7, 2018). http://dx.doi.org/10.3791/58154.

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34

Yang, Jinping, Buyun Tian, Pei Wang, et al. "SARS-CoV-2 NSP3/4 control formation of replication organelle and recruitment of RNA polymerase NSP12." Journal of Cell Biology 224, no. 3 (2024). https://doi.org/10.1083/jcb.202306101.

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β-coronavirus rearranges the host cellular membranes to form double-membrane vesicles (DMVs) via NSP3/4, which anchor replication–transcription complexes (RTCs), thereby constituting the replication organelles (ROs). However, the impact of specific domains within NSP3/4 on DMV formation and RO assembly remains largely unknown. By using cryogenic-correlated light and electron microscopy (cryo-CLEM), we discovered that the N-terminal and C-terminal domains (NTD and CTD) of SARS-CoV-2 NSP3 are essential for DMV formation. Nevertheless, the CTD of NSP4 is not essential for DMV formation but regula

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