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Berndt4 project protocol

Aging study: Pulmonary adenomas in 28 inbred strains of mice, age 20 months   (2011)

Berndt A, Harrison DE, Paigen B, Sundberg JP
With: Cario CL, Silva KA, Kennedy VE




Berndt4 Protocol

Project protocol - Contents

Workflow and sampling

Step
Procedure
Equipment
Data collected
1
 Euthanize at 20 months of age
CO2 chamber
-
2
 Fix, embed, section, and stain lung tissue
Microtome
-
3 Review and score slides Microscope with camera
Histopathology: frequency of adenomas, severity score

Equipment and supplies

  • CO2 chamber
  • Dissection kit
  • Microtome
  • Microscope with digital camera

Reagents and solutions

  • Feket's acid alcohol formalin
  • 70% ethanol
  • Paraffin
  • Hematoxylin and eosin (H&E)

Procedure: Lung tissue preparation

    1. Lungs are inflated with Feket's acid alcohol formalin and allowed to fix overnight.
    2. Lungs are stored in 70% ethanol.
    3. Lungs are trimmed longitudinally and cross-sectionally to yield a minimum of 4 lobes.
    4. Tissues are embedded in paraffin and sectioned at 6 microns.
    5. Sections are stained with H&E.

Procedure: Lung histopathology

    1. All slides are reviewed by the same experienced board-certified pathologist (JPS).
    2. For this study, diagnoses are limited to adenomas and occasional primary epithelial hyperplasias (argued to potentially represent an early stage in development of pulmonary neoplasias). Adenocarcinomas are not included in this study.
    3. Lesions are defined by their frequency -- percentage of female or male mice per strain diagnosed with one or more lesions.
    4. Lesions are characterized by severity based on tumor size and numbers per mouse (1=mild; 2=moderate; 3=severe; 4=extreme).
    5. Data are recorded and coded using the Mouse Disease Information System (MoDis; Sundberg et al., 2008; Sundberg et al., 2009).

Data collected by investigator

  • Adenoma frequency
  • Adenoma severity score for mice with tumors

MPD computed overall strain severity score (by strain, sex) to include all mice tested (mice without adenomas were considered to have a score of 0).



References

    Sundberg BA, Schofield PN, Gruenberger M, Sundberg JP. A data-capture tool for mouse pathology phenotyping. Vet Pathol. 2009 Nov;46(6):1230-40. doi: 10.1354/vp.09-VP-0002-S-FL. Epub 2009 Jul 15. PubMed 19605915   FullText

    Sundberg JP, Sundberg BA, Schofield P. Integrating mouse anatomy and pathology ontologies into a phenotyping database: tools for data capture and training. Mamm Genome. 2008 Jun;19(6):413-9. Epub 2008 Sep 17. PubMed 18797968   FullText