Rosen2 project protocol

Lateral septal nucleus morphology in 69 BXD recombinant inbred strains of mice   (2012)

Rosen GD
With: Talishinsky A

See also: Rosen2 animal documentation


Rosen2_protocol

Project protocol - Contents

Workflow and sampling

Step
Procedure performed
Equipment
Data collected
1
Mice anesthetized and perfused  
-
2
Brain removed and weighed  
-
3
Brain embedded and sectioned  
-
3
Lateral septum (LS) measured
Microscope
Lateral septum volume

Equipment and supplies

  • Computer-controlled microscope (Zeiss Axiophot, Zeiss, Melville NY)
  • Stereo Investigator (MBF Biosciences, Williston VT)

Procedure: Estimation of lateral septum volume

    1. All histologic data for this study were obtained from The Mouse Brain Library (MBL), a physical and Internet resource containing high-resolution images of histologically processed slides from over 3200 adult mouse brains.
    2. Cresyl violet stained slides are used (coronal or horizontal).
    3. Lateral septum volume is estimated using a computer-controlled microscope and software.
    4. The Cavalieri probe of Stereo Investigator is used to lay down a 250x250 micron grid; sections are examined every 150 microns.
    5. Cavalieri's method is used to compute an estimate of lateral septum volume.
    6. In cases of missing or damaged sections, a piece-wise parabolic estimation is used.
    7. Intra-observer reliability is assessed by blindly re-measuring 18 randomly chosen brains.
    8. Estimates are individually corrected for histological shrinkage by determining the previously computed ratio between the brain volume at fixation and that after processing.
    9. Estimates are individually adjusted for sex, age, epoch (Taylor strains vs. UTHSC Rww strains), and brain weight.

Definitions and calculations

  • LS: lateral septum

Data collected by investigator

  • lateral septum volume (corrected for shrinkage)
  • lateral septum volume (adjusted for sex, age, epoch (Taylor strains vs. UTHSC Rww strains), and brain weight