Slides were incubated overnight at 4°C with primary antiserum or follistatin-adsorbed primary antiserum. The next day, slides were incubated for 30 min with 1:400 dilution of biotinylated goat anti-rabbit IgG antibody (catalogue no. BA-1000; Vector Laboratories) prepared in 4% normal goat serum. The rest of the staining was carried out according to instructions provided in the Vectastain ABC kit. Slides were then dehydrated and coverslips were applied. For purposes of illustration, coverslips were removed from representative slides after densitometric quantitation, sections were hydrated, counterstained with 1:1 diluted Harris hematoxylin for 20 sec, and dehydrated; the cover-slips were then reapplied. ampicillin antibiotic
Gray-Scale Densitometric Analysis
Gray-scale densitometric analysis of follicles and corpora lutea was performed using a Carl Zeiss (Thornwood, NY) Ultraphot microscope, a high-resolution black and white microscope video camera (Hamamatsu CCD XC-77; Hamamatsu Photonics KK, Hamamatsu City, Japan) and camera control unit (C2400-60; Hamamatsu Photonics KK), and a computer-based, commercially available image analysis system (Image 1/AT; Universal Imaging Corporation, West Chester, PA). Microscope illumination, condenser position, diaphragm control, objective lens, projective lens, and video camera contrast enhancement and offset controls were standardized and remained unchanged throughout the analysis period.