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Integrated Correlative Light and Electron Microscopy for Geology

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Correlative light and electron microscopy (CLEM) the combination of fluorescence microscopy (FM) with high-resolution electron microscopy (EM). Integrated CLEM is a technique that performs both imaging modalities simultaneously in the same instrument without sample transfer.
SECOM is an integrated CLEM that is produced and designed by Delmic. This system is being used worldwide for cancer research, marine biology, neuroscience, and cell biology. However, a recent paper published in the Journal of Microscopy demonstrates a novel application for iCLEM in imaging sedimentary organic matter.
In this presentation, you can learn how iCLEM was applied to measure organic matter in geological samples.

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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Integrated Correlative Light and Electron Microscopy for Geology

  1. 1. Integration without compromise Imaging sedimentary organic matter with the integrated correlative light and electron microscope Geological materials
  2. 2. Integrated CLEM for Geology • Correlative light and electron microscopy (CLEM) is the combination of fluorescence microscopy (FM) with scanning electron microscopy (SEM) • The integrated CLEM techinique can be used to identify and characterize the organic matter in shale hydrocarbon reservoirs [1] • Correlative microscopy is useful to investigate the organic matter type and thermal regime in which organic nanoporosity forms [1] • It helps to reduce the uncertainty when estimating the undiscovered hydrocarbon resources [1] From Valentine & Hackley (2016).Correlative imaging was performed using sample transfer between two separate microscope systems: Leica DM-4000 for light microscopy and Hitachi SU-5000 FE-SEM for electron microscopy.
  3. 3. Sedimantary Organic Matter: SEM vs. FM Provides high resolution (nanometer scale) images. Organic matter visible in dark grey in Backscattered Electron (BSE) imaging. *Limitation: Cannot differentiate between different types of organic matter (e.g. kerogen versus solid bitumen) [2] Provides conclusive identification of type of organic matter [2] *Limitation: low resolution (~200 nm) Electron Microscopy (EM) Fluorescence Microscopy (FM)
  4. 4. Why use integrated CLEM? Challenges when using multiple instruments [3]: • Time-consuming for sample mapping • Difficulties in field relocation, reorientation • Sample damage during transfer Solution: Imaging sedimentary organic matter in shale simultaneously with a fluorescence microscope and a scanning electron microscope (Integrated CLEM) • Overcome these limitations • Obtain high resolution (SEM) and identification of organic matter type (FM)
  5. 5. 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
  6. 6. Sample preparation - 1 • Polished rock section glued to glass slide using cyanoacrylate glue • Section ground to thickness 20-30 µm, sputter coated with gold (~ 4 nm) and mounted in the SECOM iCLEM image with 40% FM and 60% EM contrast *Limitation: Misregistration of fluorescence image (green arrows) and electron image (red arrows and red layer margins due to the fact that the section was too thick
  7. 7. Sample preparation - 2 • Polished rock section glued to glass slide using cyanoacrylate glue • Section ground to thickness 20-30 µm • Section “wedged” by applying uneven pressure with fingertips to result in one side being polished to a vanishing edge • Sample scored and broken for Argon-milling, which removed scratches and made it even thinner near the vanishing edge • Section sputter coated with gold (~ 4 nm) and sample mounted as-is in the SECOM
  8. 8. Results: Sample preparation - 2 iCLEM image with 50% FM and 50% EM contrast Red arrows show authigenic micron- scale carbonate minerals, not visible in FM iCLEM image with 100% EM contrast Solid bitumen (marked) not visible in SEM, can be identified in FM
  9. 9. Results: Sample preparation - 2 Non-fluorescent solid bitumen and fluorescent amorphous organic matter (AOM) can be identified here (30% FM, 70% SEM) But they are indistinguishable here (100% SEM) High resolution features like sub- micron carbonate identified in EM, and different types of organic matter identified in FM
  10. 10. Reference [1] Hackley, P.C. et al (2016). Utilization of integrated correlative light and electron microscopy (iCLEM) for imaging sedimentary organic matter. Journal of Microscopy. 267: 1–13. [2] Hackley, P.C. & Cardott, B.J. (2016) Application of organic petrography in North American shale petroleum systems: a review. Int. J. Coal Geol.163: 8–51. [3] Timmermans, F.J. & Otto, C. (2015) Contributed review: review of integrated correlative light and electron microscopy. Rev. Sci. Instrum. 86: 011501. *All sample images of this presentation are extracted from the publication “Utilization of integrated correlative light and electron microscopy (iCLEM) for imaging sedimentary organic matter”
  11. 11. 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 integrated CLEM.

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