Evaluating Embryonic and Adult Mouse Tissue Using DigitallyScanned Slides

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Evaluating Embryonic and Adult Mouse Tissue Using
Digitally-Scanned Slides

• Julie F. FoleyGroup Leader, Special Techniques
  LaboratoryCellular and Molecular Pathology
  BranchNational Institute of Environmental Health
  Sciences

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RADIOLOGY
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PATHOLOGY
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(No Transcript)
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Application 1 Mouse Embryo Heart
Application

• Common phenotype in the genetically engineered
  mouse (GEM)
• Labor intensive to evaluate
• Numerous slides
• Numerous anatomic structures to evaluate
• Compare WT (normal) to GEM

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Step 1
1. Prepare Optimal Sample

• Collection Fixation
• Orientation
• Embedding
• Sectioning

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Sample Preparation
Orientation Sagittal Transverse Coronal
(Frontal)
Slide
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Sample Preparation
Sectioning 6 ?m sections stained with HE (12
serial sections - step in 100 ?m)
Sectioning - Staining Issues/Concerns
Consistent/Uniform Staining
Bilateral sectioning
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2. Digital Image
Step 2
Scanning Scanscope XT Scan at 20X Image
Scope - digital image capture (3.7X)
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Optimal Sample
The Good
Number of sections per slide Tissue
placement Uniform staining Coverslip placement
The Bad
The Ugly
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3. Create Animation
Step 3

• Adobe Photoshop CS4 Extended

• Background correction
• Scripts Image resize Image compression
  Alignment Rename files - number sequence
  Export all layers back to individual files

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e16.5 Mouse Embryo Heart Animation

• Adobe Flash Pro CS4
• Import image series
• Basic code - user controlled animation

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Application Stereology
Application 2 Stereology (Tissue Volume)Mouse
Pancreas e13.5 Whole Embryo Animation
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Estimating Total Volume of an Arbitrarily Shaped
Object
Principles and Practices of Unbiased
Stereology An Introduction for Bioscientists Peter
R. Mouton
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Pancreas Volume in e13.5 Mouse Embryo
Cut through region of interest  Select as few
as 10 sections Based on stereological
principles, estimate the volume
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Ruler Navigation Window Next Feature by
Aperio - ? Grid overlay
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Application 3 Corpora Lutea Counts Mouse Ovary
Assess implantation problem
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Summary
Using digitally-scanned slides, a series of
images can be stacked, aligned and animated to
evaluate specific anatomical structures in the
embryonic heart.
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Future
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TMALab
Future
Currently used for tissue micro arrays Stack
and align whole slide images Scan through
image stack Select and capture region of
interest
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Acknowledgements

• NIEHS - Special Techniques Laboratory
• Norris Flagler
• Tina Jones
• Elizabeth Ney
• Pat Stockton

Experimental Pathology Laboratories Beth Mahler
David Sabio