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What is Correlative Light and Electron Microscopy? - DELMIC

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Correlative light and electron microscopy (CLEM) is the combination of fluorescence microscopy (FM) and high-resolution electron microscopy (EM). Correlative microscopy has a wide variety of applications in the life sciences (cancer research, marine microbiology, neuroscience, and cell biology, etc.). This technique allows one to measure both thin sections and cultured cells.
SECOM is an integrated CLEM that is produced and designed by Delmic. Delmic offers a unique solution for simultaneous correlative light and electron microscopy.
In this presentation, we share the knowledge about the CLEM technique and point out the key advantages of using correlative microscopy in different areas.

For questions about correlative microscopy and the SECOM, please leave a comment below or visit www.delmic.com and send us a message. We will respond to your questions as soon as possible!

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What is Correlative Light and Electron Microscopy? - DELMIC

  1. 1. Integration without compromise 1 Correlative Light and Electron Microscopy on the SECOM platform
  2. 2. Fluorescence Microscopy • Provides functional information of the samples • Samples can be tagged with a dye or labeled using encoded fluorescent proteins • Fluorescent tags, multiple labels can be identified using light sources with different colors This diagram depicts the processes that occur which cause an object to emit fluorescent light of a characteristic wavelength subsequent to absorption of incident light of a lower wavelength
  3. 3. Electron Microscopy • Provides structural information of the samples • Samples can be visualized using non-specific heavy metal stains which bind to various components in the cell. Nanometer resolution is achievable because the wavelength of accelerated electrons is much shorter than that of visible light • High-resolution information and extra contrast are delivered by electron beam imaging. Scanning electron image of HeLa cell
  4. 4. Why combine fluorescence microscopy & electron microscopy? • Provides unique information that one or the other microscope alone cannot • The perfect tool for studying the complex relation between form and function in biology • For example, the functional information is visible in green (fluorescence image), and the structural information is visible in grey scale (SEM image) Correlative microscopy image of projection neurons in songbird brain. Imaging was performed using the SECOM platform (DELMIC).
  5. 5. Electron Microscopy (EM) Labelling with fluorophores - specific Functional information Resolution ~ 200 nm Labelling with heavy metals – non-specific Structural information Resolution ~ 5 nm Correlative microscopy: Combined FM and EM Correlative microscopy Fluorescence Microscopy (FM)
  6. 6. Key Advantages of integrated CLEM • Imaging of structure and function – “non-destructive” • Multi colour labelling – imaging of different functionalities • Relating morphology and performance/chemical activity – in situ • Imaging of unknown structure – overcomes limitation of fluorescence imaging • Long range labelling – enables study of sample heterogeneity • Unbiased identification of cells/features – overcomes limitation of only fluorescence/only electron imaging
  7. 7. Application Areas • Geology: Characterise different types of organic matter based on their fluorescence response and measure organic porosity • Materials Science: Investigate zeolite nanostructure (with electrons) and performance (with fluorescence) to enable rational catalyst design • Neuroscience: Study the mapping of the complete connectivity of the brain to understand the processes taking place in the context of the ultrastructure • Marine Biology: Study the relationship between form and function in microbial processes to understand the environmental impact • Live cell imaging: Study organisms in their native state by imaging living cells in solution simultaneously with light and electrons • Pathology: Accurately identify disease based on optical imaging, simultaneously confirmed by structural investigation in SEM, thereby avoiding additional imaging and sample preparation time for TEM
  8. 8. Integrated CLEM – The SECOM Electron source & lenses Sample Light microscope objective Mirror Vacuum Dichroic & emission filters Camera Light source Secondary electron detector SECOM - Integrated SEM and Fluorescence Microscope for simultaneous correlative imaging
  9. 9. Highlights of the SECOM • Automated overlay based on cathodoluminescence generation creates temporary fiducial markers, saving labour and allowing reliable interpretation of images • Avoid sample contamination/damage due to transport between microscopes • Image exactly the same features of interest simultaneously with light and electrons • Live cell imaging possible, allowing investigation of organisms/cells in their native environment using optical imaging in the SEM
  10. 10. Integration without compromise DELMIC B.V. Address: Kanaalweg 4, 2628 EB, Delft, The Netherlands Website: www.delmic.com Telephone: +31 (0)15 744 01 58 Email: info@delmic.com Please visit Delmic’s website to learn more about the unique solution for simultaneous correlative light and electron microscopy.

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