Correlative light-electron microscopy (CLEM) is the combination of an optical microscope – usually a fluorescence microscope – with an electron microscope. In an integrated CLEM system, the sample is imaged using an electron beam and an optical light path simultaneously. Traditionally, samples would be imaged using two separate microscopy modalities, potentially at different facilities and using different sample preparation methods. Integrated CLEM is thus considered to be beneficial because the methodology is quicker and easier, and it reduces the chance of changes in the sample during the process of data collection. Overlay of the two images is thus performed automatically as a result of the integration of two microscopes.[1]
This technique is used in order to obtain information at different length scales: the electron microscope provides high-resolution information down to the nano-scale, while the fluorescence microscope highlights the regions of interest. CLEM is used for various disciplines in the life sciences, including neuroscience, tissue research, and protein research.
In preparation for imaging with a fluorescence microscope, different methods can be used, such as fluorophores or dyes, immunolabeling, and genetically encoded fluorescent proteins. Different fluorescent labels can be used in order to highlight multiple regions of interest in the sample.[2] Recently Kumar et al. [3] combined FRET based molecular tension measurements[4] with cryo-electron microscopy to study how force on talin (a focal adhesion protein which directly links integrins to actin) is related to actin organization. Regions of high talin tension have highly aligned and linear filamentous actin while regions of low tension have less well-aligned actin structure.[3]
Electron microscopeedit
The electron microscope is used to obtain structural information at the nano-scale. Unlike an optical microscope, an electron microscope is able to surpass the diffraction limit of light. This is because the wavelength of accelerated electrons is much shorter than the wavelength of visible light.[5]
^"What is Correlative Light and Electron Microscopy?". 2018-04-25.
^ abKumar, Abhishek; Anderson, Karen L.; Swift, Mark F.; Hanein, Dorit; Volkmann, Niels; Schwartz, Martin A. (September 2018). "Local Tension on Talin in Focal Adhesions Correlates with F-Actin Alignment at the Nanometer Scale". Biophysical Journal. 115 (8): 1569–1579. Bibcode:2018BpJ...115.1569K. doi:10.1016/j.bpj.2018.08.045. ISSN 0006-3495. PMC6372196. PMID 30274833.
^Kumar, Abhishek; Ouyang, Mingxing; Dries, Koen Van den; McGhee, Ewan James; Tanaka, Keiichiro; Anderson, Marie D.; Groisman, Alexander; Goult, Benjamin T.; Anderson, Kurt I. (2016-05-09). "Talin tension sensor reveals novel features of focal adhesion force transmission and mechanosensitivity". J Cell Biol. 213 (3): 371–383. doi:10.1083/jcb.201510012. ISSN 0021-9525. PMC4862330. PMID 27161398.
Sueters - Di Meo, Josey; Liv, Nalan; Hoogenboom, Jacob P. (2016). "Using Advanced Correlative Microscopy to Study Complex Biological Samples". Encyclopedia of Analytical Chemistry. pp. 1–31. doi:10.1002/9780470027318.a9473. ISBN9780470027318.
Zonnevylle, A.C.; Van Tol, R.F.C.; Liv, N.; Narvaez, A.C.; Effting, A.P.J.; Kruit, P.; Hoogenboom, J.P. (2013). "Integration of a high-NA light microscope in a scanning electron microscope". Journal of Microscopy. 252 (1): 58–70. doi:10.1111/jmi.12071. ISSN 0022-2720. PMID 23889193.
April 10, 2024
correlative, light, electron, microscopy, clem, combination, optical, microscope, usually, fluorescence, microscope, with, electron, microscope, integrated, clem, system, sample, imaged, using, electron, beam, optical, light, path, simultaneously, traditionall. Correlative light electron microscopy CLEM is the combination of an optical microscope usually a fluorescence microscope with an electron microscope In an integrated CLEM system the sample is imaged using an electron beam and an optical light path simultaneously Traditionally samples would be imaged using two separate microscopy modalities potentially at different facilities and using different sample preparation methods Integrated CLEM is thus considered to be beneficial because the methodology is quicker and easier and it reduces the chance of changes in the sample during the process of data collection Overlay of the two images is thus performed automatically as a result of the integration of two microscopes 1 This technique is used in order to obtain information at different length scales the electron microscope provides high resolution information down to the nano scale while the fluorescence microscope highlights the regions of interest CLEM is used for various disciplines in the life sciences including neuroscience tissue research and protein research Contents 1 Fluorescence microscope 2 Electron microscope 3 References 4 Further readingFluorescence microscope editIn preparation for imaging with a fluorescence microscope different methods can be used such as fluorophores or dyes immunolabeling and genetically encoded fluorescent proteins Different fluorescent labels can be used in order to highlight multiple regions of interest in the sample 2 Recently Kumar et al 3 combined FRET based molecular tension measurements 4 with cryo electron microscopy to study how force on talin a focal adhesion protein which directly links integrins to actin is related to actin organization Regions of high talin tension have highly aligned and linear filamentous actin while regions of low tension have less well aligned actin structure 3 Electron microscope editThe electron microscope is used to obtain structural information at the nano scale Unlike an optical microscope an electron microscope is able to surpass the diffraction limit of light This is because the wavelength of accelerated electrons is much shorter than the wavelength of visible light 5 References edit BV DELMIC Automated Overlay Technical Note DELMIC request delmic com Retrieved 2017 02 08 What is Correlative Light and Electron Microscopy 2018 04 25 a b Kumar Abhishek Anderson Karen L Swift Mark F Hanein Dorit Volkmann Niels Schwartz Martin A September 2018 Local Tension on Talin in Focal Adhesions Correlates with F Actin Alignment at the Nanometer Scale Biophysical Journal 115 8 1569 1579 Bibcode 2018BpJ 115 1569K doi 10 1016 j bpj 2018 08 045 ISSN 0006 3495 PMC 6372196 PMID 30274833 Kumar Abhishek Ouyang Mingxing Dries Koen Van den McGhee Ewan James Tanaka Keiichiro Anderson Marie D Groisman Alexander Goult Benjamin T Anderson Kurt I 2016 05 09 Talin tension sensor reveals novel features of focal adhesion force transmission and mechanosensitivity J Cell Biol 213 3 371 383 doi 10 1083 jcb 201510012 ISSN 0021 9525 PMC 4862330 PMID 27161398 Voortman Lenard 2014 Integration without compromise Microscopy Today 22 6 30 35 doi 10 1017 S1551929514001199 S2CID 138111672 Further reading editSueters Di Meo Josey Liv Nalan Hoogenboom Jacob P 2016 Using Advanced Correlative Microscopy to Study Complex Biological Samples Encyclopedia of Analytical Chemistry pp 1 31 doi 10 1002 9780470027318 a9473 ISBN 9780470027318 Zonnevylle A C Van Tol R F C Liv N Narvaez A C Effting A P J Kruit P Hoogenboom J P 2013 Integration of a high NA light microscope in a scanning electron microscope Journal of Microscopy 252 1 58 70 doi 10 1111 jmi 12071 ISSN 0022 2720 PMID 23889193 Retrieved from https en wikipedia org w index php title Correlative light electron microscopy amp oldid 1188370528, wikipedia, wiki, book, books, library,
