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Tomographic molecular imaging and 3D quantification within adult mouse organs

Nature Methods volume 4, pages 31–33 (2007)Cite this article

Abstract

A convenient technology to quantify three-dimensional (3D) morphological features would have widespread applications in biomedical research. Based on combined improvements in sample preparation, tomographic imaging and computational processing, we present a procedure for high-resolution 3D quantification of structures within intact adult mouse organs. Using the nonobese diabetic (NOD) mouse model, we demonstrate a correlation between total islet β-cell volume and the onset of type-1 diabetes.

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Figure 1: Optimized labeling and computational background subtraction allows full 3D imaging of the intact adult mouse pancreas and the islet β-cell distribution.
Figure 2: Quantification of individual islets of Langerhans in intact pancreata.
Figure 3: Quantitative assessment of islet β-cell decline.

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Acknowledgements

We thank T. Bock and B. Hellman for helpful discussions. This work was supported by the Medical Research Council (H.M, J.S), the Wenner-Gren foundation (C.E.L), Novo Nordisk Fonden, the Swedish Diabetes Association (D.H), the Juvenile Diabetes Research Foundation (D.H, J.S, U.A), the Swedish Research Council (D.H, U.A) and the Kempe Foundation (U.A.).

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Author notes

  1. James Sharpe

    Present address: Center for Genomic Regulation, Passeig Marítim, 37-49, 08003, Barcelona, Spain

  2. Tomas Alanentalo and Amir Asayesh: These authors contributed equally to this work.

Authors and Affiliations

  1. Umeå Center for Molecular Medicine, Umeå University, Umeå, S-901 87, Sweden

    Tomas Alanentalo, Amir Asayesh, Christina E Lorén & Ulf Ahlgren

  2. MRC Human Genetics Unit, Western General Hospital, Crewe Rd., Edinburgh, EH4 2XU, UK

    Harris Morrison & James Sharpe

  3. Department of Medical and Clinical Genetics, Umeå University, Umeå, S-901 85, Sweden

    Dan Holmberg

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  1. Tomas Alanentalo
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  4. Christina E Lorén
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Contributions

T.A. developed the immunohistochemical protocol and performed immmunohistochemical analyses; A.A. performed the OPT and confocal scanning; H.M. performed OPT reconstructions and islet quantifications; C.E.L performed histological integrity controls; D.H. designed the NOD experimental setup; J.S. developed the subtraction script and supervised the project; U.A. initiated and supervised the project.

Corresponding authors

Correspondence to James Sharpe or Ulf Ahlgren.

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Competing interests

J.S. is the inventor on four patents relating to OPT technology.

Supplementary information

Supplementary Fig. 1

Optimized labeling allows visualization of large-scale specimen.

Supplementary Fig. 2

Example of variation in islet composition for a non-diabetic versus an overtly diabetic NOD mouse.

Supplementary Fig. 3

3D reconstructions of an intact mouse brain labeled for neuronal marker Isl1.

Supplementary Video 1

Iso-surface reconstruction of an adult wild type splenic pancreas. Movie showing a 3D OPT image of an intact splenic mouse pancreas (8 weeks). The reconstruction of the pancreas outline is based on the autofluorescence signal (grey) whereas the insulin expressing cells of the islets (red) are based on the signal from insulin specific antibodies. The specimen is ∼ 1.2 × 0.7 cm.

Supplementary Methods

Supplementary Protocol

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Alanentalo, T., Asayesh, A., Morrison, H. et al. Tomographic molecular imaging and 3D quantification within adult mouse organs. Nat Methods 4, 31–33 (2007). https://doi.org/10.1038/nmeth985

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