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Long‐term in vivo imaging of normal and pathological mouse spinal cord with subcellular resolution using implanted glass windows
Author(s) -
Fenrich Keith K.,
Weber Pascal,
Hocine Mélanie,
Zalc Maxime,
Rougon Geneviève,
Debarbieux Franck
Publication year - 2012
Publication title -
the journal of physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.802
H-Index - 240
eISSN - 1469-7793
pISSN - 0022-3751
DOI - 10.1113/jphysiol.2012.230532
Subject(s) - spinal cord , in vivo , axon , spinal cord injury , medicine , preclinical imaging , regeneration (biology) , pathology , pathological , neuroscience , anatomy , biology , microbiology and biotechnology
Key points• Chronic in vivo imaging of cellular interactions within the adult spinal cord with subcellular resolution is important for understanding cellular physiology and disease progression. • Previous approaches for chronic in vivo spinal cord microscopy have required surgery for each imaging session. • Here we describe a novel method for implanting glass windows over the exposed spinal cords of adult mice for repeated in vivo microscopy. • We show that the windows remain clear for many months after implantation, do not damage axons or blood vessels, and are useful for studying cellular dynamics after spinal cord injury. • Our method represents an original technical breakthrough for scientists involved in spinal cord research and in vivo imaging, and is a useful tool for studying cellular physiology and disease progression.Abstract Repeated in vivo two‐photon imaging of adult mammalian spinal cords, with subcellular resolution, would be crucial for understanding cellular mechanisms under normal and pathological conditions. Current methods are limited because they require surgery for each imaging session. Here we report a simple glass window methodology avoiding repeated surgical procedures and subsequent inflammation. We applied this strategy to follow axon integrity and the inflammatory response over months by multicolour imaging of adult transgenic mice. We found that glass windows have no significant effect on axon number or structure, cause a transient inflammatory response, and dramatically increase the throughput of in vivo spinal imaging. Moreover, we used this technique to track retraction/degeneration and regeneration of cut axons after a ‘pin‐prick’ spinal cord injury with high temporal fidelity. We showed that regenerating axons can cross an injury site within 4 days and that their terminals undergo dramatic morphological changes for weeks after injury. Overall the technique can potentially be adapted to evaluate cellular functions and therapeutic strategies in the normal and diseased spinal cord.