Vibrational Spectroscopic Imaging and Multiphoton Microscopy of Spinal Cord Injury

Spinal cord injury triggers a series of complex biochemical alterations of nervous tissue. Up to now, such cellular events could not be studied without conventional tissue staining. The development of optical, label-free imaging techniques could provide powerful monitoring tools with the potential t...

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Bibliographic Details
Published in:Analytical chemistry (Washington) 2012-10, Vol.84 (20), p.8707-8714
Main Authors: Galli, Roberta, Uckermann, Ortrud, Winterhalder, Martin J, Sitoci-Ficici, Kerim H, Geiger, Kathrin D, Koch, Edmund, Schackert, Gabriele, Zumbusch, Andreas, Steiner, Gerald, Kirsch, Matthias
Format: Article
Language:English
Subjects:
Analytical chemistry
Animals
Biochemistry
Chemistry
Exact sciences and technology
Mapping
Medical imaging
Microscopy
Microscopy, Fluorescence, Multiphoton - methods
Rats
Rats, Wistar
Spectrometric and optical methods
Spectroscopy, Fourier Transform Infrared - methods
Spectrum analysis
Spectrum Analysis, Raman - methods
Spinal Cord - pathology
Spinal cord injuries
Spinal Cord Injuries - pathology
Tissues
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Summary:Spinal cord injury triggers a series of complex biochemical alterations of nervous tissue. Up to now, such cellular events could not be studied without conventional tissue staining. The development of optical, label-free imaging techniques could provide powerful monitoring tools with the potential to be applied in vivo. In this work, we assess the ability of vibrational spectroscopy to generate contrast at molecular level between normal and altered regions in a rat model of spinal cord injury. Using tissue sections, we demonstrate that Fourier transform infrared (FT-IR) spectroscopy and spontaneous Raman spectroscopy are able to identify the lesion, the surrounding scar, and unharmed normal tissue, delivering insight into the biochemical events induced by the injury and allowing mapping of tissue degeneration. The FT-IR and Raman spectroscopic imaging provides the basis for fast multimodal nonlinear optical microscopy (coherent anti-Stokes Raman scattering, endogenous two-photon fluorescence, and second harmonic generation). The latter proves to be a fast tool for imaging of the lesion on unstained tissue samples, based on the alteration in lipid content, extracellular matrix composition, and microglia/macrophages distribution pattern. The results establish these technologies in the field of regeneration in central nervous system, with the long-term goal to extend them to intravital use, where fast and nonharmful imaging is required.
ISSN:0003-2700
1520-6882
DOI:10.1021/ac301938m