Chronic in vivo imaging in the mouse spinal cord using an implanted chamber

Department of Physics, Cornell University, Ithaca, New York, USA.
Nature Methods (Impact Factor: 25.95). 01/2012; 9(3):297-302. DOI: 10.1038/nmeth.1856
Source: PubMed

ABSTRACT Understanding and treatment of spinal cord pathology is limited in part by a lack of time-lapse in vivo imaging strategies at the cellular level. We developed a chronically implanted spinal chamber and surgical procedure suitable for time-lapse in vivo multiphoton microscopy of mouse spinal cord without the need for repeat surgical procedures. We routinely imaged mice repeatedly for more than 5 weeks postoperatively with up to ten separate imaging sessions and observed neither motor-function deficit nor neuropathology in the spinal cord as a result of chamber implantation. Using this chamber we quantified microglia and afferent axon dynamics after a laser-induced spinal cord lesion and observed massive microglia infiltration within 1 d along with a heterogeneous dieback of axon stumps. By enabling chronic imaging studies over timescales ranging from minutes to months, our method offers an ideal platform for understanding cellular dynamics in response to injury and therapeutic interventions.

Download full-text


Available from: Chris Schaffer, Aug 19, 2015
  • Source
    • "An alternative to the removal of a piece of skull (craniotomy) or vertebral column (laminectomy) is thinned bone preparations (Drew et al., 2010; Yang et al., 2010). Since 2012, the IVM field has seen the emergence of a number of methods that consist of installing a window on top of the exposed brain or spinal cord to prevent tissue degradation and to allow for repeated imaging up to several weeks following implementation (Farrar et al., 2012; Fenrich et al., 2012; Nimmerjahn, 2012; Ritsma et al., 2013). One important shortcoming of these types of windows, however, is the need to inject animals with potent anti-inflammatory drugs (e.g., dexamethasone) to reduce fibrosis and improve optical clarity. "
    NEUROINFLAMMATION: NEW INSIGHTS INTO BENEFICIAL AND DETRIMENTAL FUNCTIONS, 2015 edited by Samuel David, 05/2015: chapter In Vivo Imaging of Glial and Immune Cell Responses in Central Nervous System Injury and Disease: pages 21-38; John Wiley & Sons, Inc..
  • Source
    • "To overcome this downside, a custom-made imaging chamber has been proposed [18]. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Intravital microscopy has emerged in the recent decade as an indispensible imaging modality for the study of the micro-dynamics of biological processes in live animals. Technical advancements in imaging techniques and hardware components, combined with the development of novel targeted probes and new mice models, have enabled us to address long-standing questions in several biology areas such as oncology, cell biology, immunology and neuroscience. As the instrument resolution has increased, physiological motion activities have become a major obstacle that prevents imaging live animals at resolutions analogue to the ones obtained in vitro. Motion compensation techniques aim at reducing this gap and can effectively increase the in vivo resolution. This paper provides a technical review of some of the latest developments in motion compensation methods, providing organ specific solutions.
    IEEE Journal of Selected Topics in Quantum Electronics 03/2014; 20(2). DOI:10.1109/JSTQE.2013.2279314 · 3.47 Impact Factor
  • Source
    • "These observations are consistent with the hypothesis that regenerating axons form synapse-like terminals with reactive glia, which was originally advanced by Carlstedt (1985). Advances in imaging have allowed live in vivo studies of acute and chronically injured axons in the lesion environment (Di Maio et al., 2011; Evans et al., 2014; Farrar et al., 2012; Kerschensteiner et al., 2005; Ylera et al., 2009). Ylera et al. (2009) recently used in vivo imaging to demonstrate that chronically injured axons can, indeed, be aroused into a robust regenerative state. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Astrocytes react to CNS injury by building a dense wall of filamentous processes around the lesion. Stromal cells quickly take up residence in the lesion core and synthesize connective tissue elements that contribute to fibrosis. Oligodendrocyte precursor cells proliferate within the lesion and help to entrap dystrophic axon tips. Here we review evidence that this aggregate scar acts as the major barrier to regeneration of axons after injury. We also consider several exciting new interventions that allow axons to regenerate beyond the glial scar, and discuss the implications of this work for the future of regeneration biology.
    Experimental Neurology 01/2014; 253. DOI:10.1016/j.expneurol.2013.12.024 · 4.62 Impact Factor
Show more