Comparison of Plaque Burden in Tg2576 Mice Using the Aperio and TurboScan Imaging Platforms

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Comparison of Plaque Burden in Tg2576 Mice Using the Aperio and TurboScan Imaging Platforms A.J. Milici. Alzheimer’s Disease. Alzheimer’s Disease (AD) is a progressive neurodegenerative disorder of the brain and is the most common form of dementia. - PowerPoint PPT Presentation

Text of Comparison of Plaque Burden in Tg2576 Mice Using the Aperio and TurboScan Imaging Platforms

  • Comparison of Plaque Burden in Tg2576 Mice Using the Aperio and TurboScan Imaging Platforms

    A.J. Milici

  • Alzheimers DiseaseAlzheimers Disease (AD) is a progressive neurodegenerative disorder of the brain and is the most common form of dementia.Major contributing factor for increased risk is old age.After 65 a doubling of risk occurs every 5 years effecting ~30% of people >80 years old.Two histopathological changes within the brain.Neurofibrillary TanglesAmyloid Plaques

  • Amyloid PlaqueAmyloid plaque occurs extracellular to the neuron cell body and is diagnostic of Alzheimers disease (AD).Plaque made up of a 42 amino acid b amyloid peptide (Ab42).Ab42 originates from abnormal processing of Amyloid Precursor Protein (APP).APP is a transmembrane cell surface protein with functional roles in:Neurite outgrowthCell adhesionSynaptic functionsInduction of apoptosis

  • What has been previously done to quantify Amyloid?Many previous studies have sampled a few fields from cortex and hippocampusIssuesVariability of transgene expressionSmall N Selective field sampling bias

  • MethodsHemibrains were fixed ON in NBFRoutinely processed and embedded in Paraffin15 m serial sections were collectedSections were routinely stained with 6E10 antibodyImages were captured:@10x using TurboScan on a Nikon--- microscope@20x using an AperioXT systemTurboScan images were analyzed using ImageProAperio images were analyzed using the Positive Pixel Algorithm

  • Can we easily capture and analyze whole brain sections?Chose to analyze sagittal sectionsAllowed comparison of histological data to biochemical data from the same animal for both cortex and hippocampusFirst key question was how many sections/animal need to be analyzedInitially requested to serially section and stain an entire brain.

  • How many sections do we need to analyze?Able to demonstrate that 2-3 sections (15 m thick) evenly spaced resulted in a similar dataset obtained with 8 sections.This correlation held up with animals as they aged and developed more plaque.

  • Collect image montage @10x using TurboScanSagittal section stained with 6E10Mosaic made up of 60-70 images ~260 Mb File size creates ImagePro issuesRequires additional pre-image processing steps to reduce image size to
  • Invert contrast to enhance discrimination of plaque

  • Draw AOI around Cortex

  • Threshold and measure objects > 25 um2

  • Objects in Hippocampus

  • Excel Data Output

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