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Correlative light and electron microscopy for type 1 diabetes

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Correlative light and electron microscopy (CLEM) is the combination of fluorescence microscopy (FM) and high-resolution electron microscopy (EM). Diabetes Type 1, one of the two widely spread forms, is an autoimmune decease, which is caused by the destruction of insulin-producing beta cells in the pancreas. A better understanding of this form of Diabetes could be obtained from a rigorous study of the islets of Langerhans. This can be done by imaging them with an electron microscope (EM) and fluorescence microscope combined in one. This technique called correlative microscopy is commonly applied in life sciences.
The SECOM platform 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, you can learn how integrated CLEM can be performed to study and better understand type 1 diabetes.
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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Correlative light and electron microscopy for type 1 diabetes

  1. 1. Integration without compromise 1 Correlative Microscopy for understanding Diabetes Type 1
  2. 2. Type 1 Diabetes • Type 1 diabetes occur when the insulin-producing beta cells are attacked and destroyed. This could potentially lead to cell death from lack of glucose and multiple organ failure due to sustained high blood sugar levels. • It is estimated that 9% of the population is affected by this life-threatening decease [1]. [1] G. Danaei et al., The Lancet 378, 9785, 31-40 (2011) Source: Google Sites
  3. 3. Electron Microscopy Advantages • High spatial resolution • Allows the identification of subcellular features Limitations • Slow and highly labor-intensive • Hard to identify specific cells/features in a grey-scale image Using electron microscopy in diabetes type 1 research
  4. 4. Fluorescence Microscopy Using fluorescence microscopy in diabetes type 1 research Advantages • Allows multicolor imaging • Identification of cells possible over a large field of view Limitations • Resolution is diffraction limited (~200nm) • Unable to provide structural information of the samples
  5. 5. Correlative Light and Electron Microscopy for Diabetes Type 1 • Integrated correlative light and electron microscopy (iCLEM) is the combination of fluorescence microscopy (FM) with high-resolution electron microscopy (EM) in one • iCLEM offers a powerful imaging technique where by the cells of interest can be identified magnification EM image can be acquired at the desired location to reveal structural details • This can be used to image the Islets of Langerhans for type 1 diabetes research [2].
  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. 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
  8. 8. iCLEM of rat pancreas Sample preparation • 80 nm thick sections of healthy rat pancreas were prepared for correlative • Fresh pancreas were fixed in 4% para - formaldehyde and 0.1% selected for the presence of islets of Langerhans. • Post-fixation was carried out with 1% osmium tetroxide, followed by EPON. • Ultrathin sections were then cut and mounted on ITO-coated glass slides. labelling was performed with three different fluorophores. • The sample was imaged on the SECOM platform using an automated Results As seen in the image, the labelling of the insulin (beta cells) in orange with visible and the ultrastructure can be examined in detail from the EM contrast. guanine quadruplexes and the nucleus. These results demonstrate the diabetes type 1 [2]. Sample courtesy: B.N.G. Giepmans & P. de Boer, UMC G
  9. 9. References [1] G. Danaei et al., The Lancet 378, 9785, 31-40 (2011) [2] J. Kuipers et al., Experimental Cell Research 337, 202–207 (2015)
  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 correlative light and electron microscopy.