Scion lg 3 frame grabber

DAB-stained sections were gently placed onto gel-coated slides, dehydrated in alcohol and Citrisolv (Fisher), then coverslipped and left to dry. A light microscope (Leica, DMR) and a Swanson rat brain atlas (Swanson, 2004, Brain Maps: Structure of Rat Brain) were used to identify the sections. As previously described in detail [21 (link)], images of the BNSTov, CEAl, PVN, DMH, Pi and SCN were captured with a Sony XC-77 video camera, Scion LG-3 frame grabber (Scion Corporation, Frederick, MD, USA), and Image SXM software (http://www.ImageSXM.org.uk1 v1.95, S.D. Barret). Using counts from six unilateral images showing the highest number of labeled nuclei, the mean number of PER1 immunoreactive (IR) cells per region was then calculated for each animal, as previously described [13 (link),21 (link)]. Capturing of images and counting of cells were performed blind. Data were analyzed using a two-way analysis of variance (ANOVA), with a confidence level [α] set at 0.05. Differences between stressed and control groups were revealed using Bonferroni or Dunnett’s post-hoc analyses. In both the ADX and GRX experiments, data from control groups for restraint and 2DG were tested and determined to have no significant differences (data not shown), and accordingly, pooled together.

Stained sections were mounted onto gel-coated slides and dehydrated in a series of alcohols and Citrisolv (Fisher), then coverslipped. The sections were examined under a light microscope (Leica, DMR) and identified using a Swanson rat brain atlas [40] . Images of the SCN, PVN, DMH, Pi, BNSTov and CEA were captured using a Sony XC-77 video camera, Scion LG-3 frame grabber (Scion Corporation, Frederick, MD, USA), and Image SXM software (S.D. Barret, v1.95, http://www.ImageSXM.org.uk). The mean number of PER1 and FOS IR cells per region was then calculated for each animal from the counts of six unilateral images showing the highest number of labeled nuclei, as previously described [9] (link), [10] (link). All images were sampled from the entire nucleus in question, at different rostral-caudal levels. Image capture and cell counts were performed blind. Data were analyzed using a two-way analysis of variance (ANOVA) and a Bonferroni post-hoc test. Differences between groups for the time of day effect of stress were revealed using a one-way ANOVA and Dunnett's post-hoc analysis.