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Book Chapter

Intravital Imaging of Autoreactive T Cells in Living Animals

MPS-Authors
http://pubman.mpdl.mpg.de/cone/persons/resource/persons38918

Kawakami,  Naoto
Emeritus Group: Neuroimmunology / Wekerle, MPI of Neurobiology, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons38757

Bartholomäus,  Ingo
Emeritus Group: Neuroimmunology / Wekerle, MPI of Neurobiology, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons61113

Pesic,  Marija
Emeritus Group: Neuroimmunology / Wekerle, MPI of Neurobiology, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons98405

Kyratsous,  Nikolaos I.
Emeritus Group: Neuroimmunology / Wekerle, MPI of Neurobiology, Max Planck Society;

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Citation

Kawakami, N., Bartholomäus, I., Pesic, M., & Kyratsous, N. I. (2012). Intravital Imaging of Autoreactive T Cells in Living Animals. In LABORATORY METHODS IN CELL BIOLOGY: IMAGING (pp. 149-168). 525 B STREET, SUITE 1900, SAN DIEGO, CA 92101-4495 USA: ELSEVIER ACADEMIC PRESS INC.


Cite as: http://hdl.handle.net/11858/00-001M-0000-0013-FB80-0
Abstract
Developing technologies now allow observing cellular motility and functions in the living animal. Here, we introduce the method of intravital imaging by using two-photon microscopy. To acquire images with good quality, a highly stablilized tissue is required. Additionally, physiological parameters of the animal need to be controlled during the entire period of intravital imaging. We image rat autoreactive T cells within the spinal cord leptomeninges. Those autoantigen specific T cells were labeled in vitro by using fluorescent protein coding retroviral vectors. Adoptively transferred T cells migrate into the spinal cord with highly reproducible time kinetic. Intravital imaging was performed within the deeply anesthetized animals. Although two-photon microscopy is a powerful tool, the penetration depth in certain tissues, like the spinal cord parenchyma, is still limited. To overcome this issue, imaging of explanted acute spinal cord slices was performed under nearly physiological conditions. Results obtained from intravital imaging will strengthen the "snap shots" analysis such as FACS and quantitative PCR, and can provide new insight into cellular mechanisms in vivo.