Brain Histopathology MGP_BHP_001

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Purpose

To perform a histopathological examination of the brain (striatal, hippocampal and/or cerebellar sections) and to record abnormal findings.

To perform a histopathology examination and annotation on a standard list of parameters of the brain (striatal, hippocampal and/or cerebellar sections).

Experimental Design

Number of mutant animals: 2-3 mutant males

Typical age at experiment 6-16 weeks. Mutants are compared to wildtype mice of the same age

Procedure

  1. Collect and weigh the brains.
  2. Drop fix in PFA or 10% Neutral Buffered Formalin and ship weekly.
  3. Trim the brain producing 3 chunks per brain for the 3 brain areas.
  4. Place each brain area on its corresponding cassette and incubate at room temperature in 70% alcohol.
  5. Place cassettes onto tissue processor for 18hr 30min for dehydration with alcohol, replacement with solvent and incubation with paraffin.
  6. Properly embed the brain chunks in paraffin onto the sectioning cassettes.
  7. Section each brain area to obtain the 4 critical 5um sections per area to be placed on slides.
  8. Store the slides in a slide box without a lid to dry overnight at 37oC in an incubator.
  9. Double stain the slides with nissl-luxol and coverslip.
  10. Select the best critical section for each area and scan on a Hamamatsu.
  11. Analyse and record the standard list of parameters for each brain area.
  12. QC data.
  13. Deblind, group by genotype and perform statistical analysis.
  14. Upload and annotate images in Centre-specific database using the standardized IMPC Gross Pathology ontology:
    1. MA ontology terms for location/topography
    2. MP ontology terms for large/small/abnormal
  15. Record all abnormal findings in a Center-specific database using the standardized IMPC Gross Pathology ontology:
    1. MA ontology terms for location/topography
    2. MP ontology terms for large/small/abnormal

Parameters & Metadata

Filter:

Name Version Type Description Req. Upload Req. Analysis Annotation Increment Options Ontology Options Unit Data Type Derived History Key Group ID
Name Version Type Description Req. Upload Req. Analysis Annotation Increment Option Ontology Options Unit Data Type Derived History Key Group ID