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High-throughput quantitative histological analysis of Alzheimer's disease pathology using a confocal digital microscanner

Nature Biotechnology volume 17, pages 53–57 (1999)Cite this article

Abstract

To develop a rapid method of quantifying immunohistochemical information in tissue sections, we tested a confocal laser fluorescence microscanner initially designed for DNA microarray analysis. This instrument collects digital images at multiple wavelengths, scans entire sections at a resolution of 5 or 10 μm in less than 10 min, and quantifies structures labeled with fluorescent or nonfluorescent probes. We assessed the microscanner by studying immunostained amyloid plaques in the Alzheimer's disease (AD) brain and in the brain of a transgenic mouse model of AD amyloidosis, as efforts to correlate measures of amyloid plaques in brain sections with behavioral impairments are impeded by limitations in current morphometric methods. Microscanner analysis was used to determine amyloid burden in the occipital and entorhinal cortices of the mouse (3.7%) and human AD brain (1.6%). We also quantified the colocalization of plaque b-amyloid (Ab) with glial fibrillary acidic protein, a marker of gliosis (mouse 0.9%, human AD 3.7%). The microscanner may be generally applicable to a wide variety of human histopathologies and their animal models, wherever rapid unbiased quantitative analysis is needed.

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Figure 1: Comparison of the field uniformity of (A) a confocal microscope and (B) the microscanner.
Figure 2: Transgenic mouse brain sections stained for Aβ with antibody 2332 and Cy5 (A) or Cy3 (B).
Figure 3: (A) Section of human cortex from an AD patient.
Figure 4: Amyloid burden in entorhinal/occipital cortex of PDAPP mouse.
Figure 5: Distribution of Aβ and GFAP immunoreactivity in mouse and human sections.
Figure 6: Comparison of feature extraction methods.

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Acknowledgements

We thank V.M.-Y. Lee, D.L. Barker, Y. Nakagawa, K.B. Bechtol, and M.L. Schmidt as well as T.-H. Chiu, M.T. Schuck, and B. Kain for advice and assistance with these studies. Appreciation also is expressed to J. Clemens and members of his laboratory at Eli Lilly for making the PDAPP mice available to us for this study, and to the families of the patients studied here who made this research possible through their generosity and support. Supported by grants from the National Institute on Aging of the National Institutes of Health to J.Q.T., and by an ATP/NIST grant (70NANB5H1031) to Molecular Dynamics.

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Authors and Affiliations

  1. Molecular Dynamics, Inc., Sunnyvale, 94086, CA

    David K. Hanzel, Richard F. Johnston & Jeanne F. Loring

  2. Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, 19104, PA

    John Q. Trojanowski

  3. ARC Genomic Research, Foster City, 94404, CA

    Jeanne F. Loring

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  1. David K. Hanzel
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  2. John Q. Trojanowski
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  4. Jeanne F. Loring
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Correspondence to David K. Hanzel.

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Hanzel, D., Trojanowski, J., Johnston, R. et al. High-throughput quantitative histological analysis of Alzheimer's disease pathology using a confocal digital microscanner. Nat Biotechnol 17, 53–57 (1999). https://doi.org/10.1038/5225

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