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.