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Bright-Field Imaging and Optical Coherence Tomography of the Mouse Posterior Eye

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Mouse Models for Drug Discovery

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1438))

Abstract

Noninvasive live imaging has been used extensively for ocular phenotyping in mouse vision research. Bright-field imaging and optical coherence tomography (OCT) are two methods that are particularly useful for assessing the posterior mouse eye (fundus), including the retina, retinal pigment epithelium, and choroid, and are widely applied due to the commercial availability of sophisticated instruments and software. Here, we provide a guide to using these approaches with an emphasis on post-acquisition image processing using Fiji, a bundled version of the Java-based public domain software ImageJ. A bright-field fundus imaging protocol is described for acquisition of multi-frame videos, followed by image registration to reduce motion artifacts, averaging to reduce noise, shading correction to compensate for uneven illumination, filtering to improve image detail, and rotation to adjust orientation. An OCT imaging protocol is described for acquiring replicate volume scans, with subsequent registration and averaging to yield three-dimensional datasets that show reduced motion artifacts and enhanced detail. The Fiji algorithms used in these protocols are designed for batch processing and are freely available. The image acquisition and processing approaches described here may facilitate quantitative phenotyping of the mouse eye in drug discovery, mutagenesis screening, and the functional cataloging of mouse genes by individual laboratories and large-scale projects, such as the Knockout Mouse Phenotyping Project and International Mouse Phenotyping Consortium.

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Authors and Affiliations

  1. The Jackson Laboratory, 600 Main Street, Bar Harbor, ME, 04609, USA

    Mark P. Krebs Ph.D., Mei Xiao, Keith Sheppard, Wanda Hicks & Patsy M. Nishina

Authors
  1. Mark P. Krebs Ph.D.
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  2. Mei Xiao
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  3. Keith Sheppard
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  4. Wanda Hicks
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  5. Patsy M. Nishina
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Corresponding author

Correspondence to Mark P. Krebs Ph.D. .

Editor information

Editors and Affiliations

  1. Takeda Pharmaceutical Company, Cambridge, Massachusetts, USA

    Gabriele Proetzel

  2. The Jackson Laboratory, Bar Harbor, Maine, USA

    Michael V. Wiles

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© 2016 Springer Science+Business Media New York

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Krebs, M.P., Xiao, M., Sheppard, K., Hicks, W., Nishina, P.M. (2016). Bright-Field Imaging and Optical Coherence Tomography of the Mouse Posterior Eye. In: Proetzel, G., Wiles, M. (eds) Mouse Models for Drug Discovery. Methods in Molecular Biology, vol 1438. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3661-8_20

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  • DOI: https://doi.org/10.1007/978-1-4939-3661-8_20

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3659-5

  • Online ISBN: 978-1-4939-3661-8

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