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In Vivo Visualization of (Auto)Immune Processes in the Central Nervous System of Rodents

  • Christian Schläger
  • Tanja Litke
  • Alexander Flügel
  • Francesca Odoardi
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1304)

Abstract

The CNS is effectively shielded from the periphery by the blood–brain barrier (BBB) which limits the entry of cells and solutes. However, in autoimmune disorders such as multiple sclerosis, immune cells can overcome this barrier and induce the formation of CNS inflammatory lesions. Recently, two-photon laser scanning microscopy (TPLSM) has made it possible to visualize autoimmune processes in the living CNS in real time. However, along with a high microscopy standard, this technique requires an advanced surgical procedure to access the region of interest. Here, we describe in detail the necessary methodological steps to visualize (auto)immune processes in living rodent tissue. We focus on the procedures to image the leptomeningeal vessels of the thoracic spinal cord during transfer experimental autoimmune encephalomyelitis in LEW rats (AT EAE) and in active EAE in C57BL/6 mice (aEAE).

Keywords

Dura (mater) spinalis Experimental autoimmune encephalomyelitis (EAE) Encephalitogenic T cells Laminectomy TPLSM (two-photon laser scanning microscopy) Spinal cord window myelin basic protein (MBP) myelin oligodendrocyte glycoprotein (MOG) 

Notes

Acknowledgments

The authors thank C. Ludwig for text editing. We thank N. Kawakami and I. Bartholomðus for their help in setting up the TPLSM technique and surgical procedures. This work was supported by the Deutsche Forschungsgemeinschaft (TRR-SFB43 projects B10 and B11, FORR 1336), the Bundesministerium für Bildung und Forschung (“UNDERSTAND MS”), and the Ministry of Science and Culture of Lower Saxony (Niedersachsen-Research Network on Neuroinfectiology, N-RENNT).

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Christian Schläger
    • 1
  • Tanja Litke
    • 1
  • Alexander Flügel
    • 1
    • 2
  • Francesca Odoardi
    • 1
    • 2
  1. 1.Department of Neuroimmunology, Institute for Multiple Sclerosis ResearchUniversity Medical Centre Göttingen and Gemeinnützige Hertie-StiftungGöttingenGermany
  2. 2.Max-Planck-Institute for Experimental MedicineGöttingenGermany

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