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Title: Evaluating Embryonic and Adult Mouse Tissue Using DigitallyScanned Slides


1
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

2
RADIOLOGY
3
PATHOLOGY
4
(No Transcript)
5
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

6
Step 1
1. Prepare Optimal Sample
  • Collection Fixation
  • Orientation
  • Embedding
  • Sectioning

7
Sample Preparation
Orientation Sagittal Transverse Coronal
(Frontal)
Slide
8
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
9
2. Digital Image
Step 2
Scanning Scanscope XT Scan at 20X Image
Scope - digital image capture (3.7X)
10
Optimal Sample
The Good
Number of sections per slide Tissue
placement Uniform staining Coverslip placement
The Bad
The Ugly
11
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

12
e16.5 Mouse Embryo Heart Animation
  • Adobe Flash Pro CS4
  • Import image series
  • Basic code - user controlled animation

13
Application Stereology
Application 2 Stereology (Tissue Volume)Mouse
Pancreas e13.5 Whole Embryo Animation
14
Estimating Total Volume of an Arbitrarily Shaped
Object
Principles and Practices of Unbiased
Stereology An Introduction for Bioscientists Peter
R. Mouton
15
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
16
Ruler Navigation Window Next Feature by
Aperio - ? Grid overlay
17
Application 3 Corpora Lutea Counts Mouse Ovary
Assess implantation problem
18
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
21
Acknowledgements
  • NIEHS - Special Techniques Laboratory
  • Norris Flagler
  • Tina Jones
  • Elizabeth Ney
  • Pat Stockton

Experimental Pathology Laboratories Beth Mahler
David Sabio
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