1. Morphometric and ultrastructural analysis of human granulosa cells after gonadotrophin- releasing hormone agonist or antagonist 
L. Centurione, F. Giampietro, S. Sancilio, M. Piccirilli, L. Artese, G.M. Tiboni, R. Di Pietro 
Reproductive BioMedicine Online 
Volume 20, Issue 5, Pages (May 2010)
DOI: /j.rbmo
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2. Figure 1
Light microscopy identification of human granulosa cell subpopulations from GnRH agonist- or antagonist-treated women on toluidine blue-stained semithin sections. Note morphological differences between pale (arrowhead) and dark (arrow) cells. Magnification ×100.
Reproductive BioMedicine Online  , DOI: ( /j.rbmo )
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3. Figure 2
Distribution of human granulosa cell subpopulations derived from GnRH agonist- or antagonist-treated women. Values are expressed as mean percentage±SEM. White bars=pale cells; black bars=dark cells; ∗P<0.05.
Reproductive BioMedicine Online  , DOI: ( /j.rbmo )
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4. Figure 3
Threshold method employed to analyse different cell parameters using MetaMorph image analysis software on toluidine blue-stained semithin sections. Acquired images were analysed to evaluate cell area of (a) pale cells and (b) dark cells or (c) to measure the size of each toluidine blue-positive granule. Red fields created by the software were considered for the analysis. Note the different colour intensity of granules in the pale and dark cells. Magnification ×100.
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5. Figure 4
Box plots demonstrating the central tendency of the values, their variability, the symmetry of the distribution and the presence of outliers (values out of the significant range). (a) Cell area (μm2); (b) granule/cell area; (c) granule size (μm2). The lower edge of the box represents the first quartile; the black line represents the median, the red line represents the average. The upper edge of the box represents the third quartile. Circles are filled when the values are more than three times the distance between the first and third quartiles. Circles are open if they are within that interval.
Reproductive BioMedicine Online  , DOI: ( /j.rbmo )
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6. Figure 5
Representative electron micrographs showing ultrastructural details of pale and dark granulosa cells. (a) The large pale cell was identified as a large cell with a typical round and euchromatic nucleus and pale cytoplasm (×7000; inset shows the pale cell at magnification ×3000), mainly due to the presence of an abundant SER (a1: arrows, ×30,000). In close relationship with these structures, a number of osmiophilic granules identified as lipid droplets of various size and electron-density (a2: arrowheads, ×30,000) as well as of polymorphic mitochondria were also observed. At the nuclear level, the presence of numerous active nuclear pores (a3; stars, ×30,000) was clearly evident. (b) The small/dark cell was identified as a shrunken cell with an indented and heterochromatic nucleus and nuclear pores rarely evident (×7000, inset shows the dark cell at magnification ×3000). In the dark cytoplasm, due to the presence of numerous free ribosomes and scarce endoplasmic reticulum, only a few mitochondria were evident (b1; M arrow, ×30,000), displaying condensed cristae and not in direct contact with reticulum or lipid droplets. These droplets appeared large, dark and with a homogeneous osmiophilic content of lipid nature and, interestingly, displayed aspects of degeneration presumably attributable to the numerous lysosomes that were closely associated with them (b2; L arrow, ×30,000), as seen at a higher magnification (b3, ×50,000).
Reproductive BioMedicine Online  , DOI: ( /j.rbmo )
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