Uteri were dissected and the uterine wet weights were measured with analytical
electron balancer. A part of uterus was stored at −80° until use
for molecular experiments and the others were fixed overnight in Bouin's
fixative. Fixed uteri were dehydrated with ethyl alcohol using Leica TP 1020,
and embedded in paraffin. The paraffin block was cross-sectioned at 4 μm
(Leica RM2245 microtome) and stained with hematoxylin and eosin Y. Tissues are
microphotographed using Olympus B×60 microscope and Olympus DP71
microscope digital camera. Uterine diameter was measured perpendicularly to
mesometrium-antimesometrium axis on 40× microphotograph with ImageJ
program. Myometrium and endometrium thickness were respectively measured from
longitudinal to circular smooth muscle layers and from luminal surface to
beginning of circular smooth muscle layer on 100× microphotograph with
ImageJ software. Epithelial cell height was measured on 400×
magnification microscope with Tcapture software. Furthermore, number of
endometrial glands was counted and its morphology was analyzed. To get
confidence of the data, at least 4 sections per mouse were analyzed and in all
directions.
electron balancer. A part of uterus was stored at −80° until use
for molecular experiments and the others were fixed overnight in Bouin's
fixative. Fixed uteri were dehydrated with ethyl alcohol using Leica TP 1020,
and embedded in paraffin. The paraffin block was cross-sectioned at 4 μm
(Leica RM2245 microtome) and stained with hematoxylin and eosin Y. Tissues are
microphotographed using Olympus B×60 microscope and Olympus DP71
microscope digital camera. Uterine diameter was measured perpendicularly to
mesometrium-antimesometrium axis on 40× microphotograph with ImageJ
program. Myometrium and endometrium thickness were respectively measured from
longitudinal to circular smooth muscle layers and from luminal surface to
beginning of circular smooth muscle layer on 100× microphotograph with
ImageJ software. Epithelial cell height was measured on 400×
magnification microscope with Tcapture software. Furthermore, number of
endometrial glands was counted and its morphology was analyzed. To get
confidence of the data, at least 4 sections per mouse were analyzed and in all
directions.