Measuring area and intensity of growing follicles located in thick ovarian tissue
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On the right are confocal microscope images of solid ovary tissue derived with a Leica TCS4d microscope. The specimen was fixed with paraformaldehyde, stained with YoPro, dehydrated with MEOH and cleared with benzyl alcohol/ benzyl benzoate. The individual sections of the ovarian follicle were obtained at the focus of the nucleolus. The entire follicle was taken with a 10 x objective (Mag 100x) while the individual oocyte contained indside the follicle was taken with a 20 x objective ( zoom 3, Mag 600x) the individual components of the follicle was measured.
Image-Pro Plus was used to measure area and intensity of growing follicles located in thick ovarian tissue. The area and intensity of the follicle, nucleus, nucleolus, oocyte, Zona pellucida, and cumulus cells were obtained. If necessary the gamma/brightness/contrast were varied to reveal structures in the oocyte that had reduced gray scale values.
Submitted by:
Dr. Robert Zucker
USEPA
NHEERL
Reproductive Toxicology Division
Research Triangle Park, North Carolina 27711
zucker.robert@epamail.epa.gov
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