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Orthogonal polarization spectral imaging: A new method for study of the microcirculation

Nature Medicine volume 5, pages 1209–1212 (1999)Cite this article

Different disease states, including diabetes, hypertension and coronary heart disease, produce distinctive microvascular pathologies. So far, imaging of the human microcirculation has been limited to vascular beds in which the vessels are visible and close to the surface (for example, nailfold, conjunctiva). We report here on orthogonal polarization spectral (OPS) imaging, a new method for imaging the microcirculation using reflected light that allows imaging of the microcirculation noninvasively through mucus membranes and on the surface of solid organs. In OPS imaging, the tissue is illuminated with linearly polarized light and imaged through a polarizer oriented orthogonal to the plane of the illuminating light. Only depolarized photons scattered in the tissue contribute to the image. The optical response of OPS imaging is linear and can be used for reflection spectrophotometry over the wide range of optical density typically achieved by transmission spectrophotometry. A comparison of fluorescence intravital microscopy with OPS imaging in the hamster demonstrated equivalence in measured physiological parameters under control conditions and after ischemic injury. OPS imaging produced high-contrast microvascular images in people from sublingual sites and the brain surface that appear as in transillumination. The technology can be implemented in a small optical probe, providing a convenient method for intravital microscopy on otherwise inaccessible sites and organs in the awake subject or during surgery for research and for clinical diagnostic applications.

At present, the use of microvascular imaging in diagnosis and treatment of human disease is limited. Use has been made of nailfold capillaroscopy in the diagnosis and treatment of peripheral vascular diseases, diabetes and hematological disorders1,2,3. Problems with movement have restricted the use of the bulbar conjunctiva for clinical applications in opthalmology4,5,6. Other locations observed by intravital microscopy include the microcirculation of the skin, lip, gingival tissue and tongue4. Laser-scanning confocal imaging is one new technique that does allow reflected light imaging of the microcirculation in vivo 7,8. This method can distinguish between layers of skin and can image microcirculation in the skin. However, images obtained using laser-scanning confocal microscopy can only be collected at a fraction of the normal video rate (up to 16 versus 30 frames/s) and the best microvascular images using this technique require fluorescent labels for contrast enhancement7. Direct observation, using conventional methods, of vascular beds of other organs in humans has been prevented by the need for transillumination, fluorescent dyes for contrast enhancement, or by the size of instrumentation required to acquire images, especially during surgery.

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Figure 1: a, Orthogonal polarization spectral imaging probe.
Figure 2: Images of the microcirculation dorsal skinfold preparation of the awake hamster taken during fluorescence intravital microscopy (a and c) and OPS imaging (b and d).
Figure 3: Quantitative determination of optical density in vitro.
Figure 4: OPS images of human microcirculation.

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Acknowledgements

We thank A. Loeb, G. Fletcher, C. Cook, A. Perkins, R. Pittman and R. Johnson for discussions and critically reviewing the manuscript. These studies were funded in part by Cytometrics.

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Authors and Affiliations

  1. Cytometrics, Inc., 615 Chestnut Street , Philadelphia, 19106, Pennsylvania, USA

    Warren Groner & Richard G. Nadeau

  2. Brigham and Women's Hospital, Harvard Medical School , 75 Francis Street, Boston, 02115, Massachusetts , USA

    James W. Winkelman

  3. Institute for Surgical Research, Klinikum Grosshadern, University of Munich, Marchioninistrasse 15, Munich, 81377, Germany

    Anthony G. Harris & Konrad Messmer

  4. Department of Anesthesiology, Academic Medical Center University of Amsterdam, Meibergdreef 9, Amsterdam, 1105 AZ, The Netherlands

    Can Ince

  5. Department of Neurosurgery, Academic Medical Center University of Amsterdam, Meibergdreef 9, Amsterdam, 1105 AZ, The Netherlands

    Gerrit J. Bouma

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  1. Warren Groner
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Correspondence to Richard G. Nadeau.

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Groner, W., Winkelman, J., Harris, A. et al. Orthogonal polarization spectral imaging: A new method for study of the microcirculation. Nat Med 5, 1209–1212 (1999). https://doi.org/10.1038/13529

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