Diffraction-enhanced Synchrotron Imaging of Bovine Ovaries Ex Vivo

The objective of this study was to test the hypothesis that diffraction-enhanced imaging (DEI), a synchrotron x-ray imaging technique, would provide greater contrast for evaluating bovine ovaries compared with conventional diagnostic ultrasonography.

Bovine ovaries were evaluated ex vivo as follows: fresh without radiographic arterial contrast (n = 2), fresh with contrast (n = 1), preserved in 10% formalin without contrast (n = 2), and preserved with contrast (n = 1). Each ovary was imaged with DEI and subsequently with ultrasonography and histology. The ability to visualize and differentiate preantral and antral follicles, corpora lutea (CL), and cumulus oocyte complexes (COCs) were compared using DEI, ultrasonography, and histology. The diameter of follicles and CL were measured and compared using ultrasonography, DEI, and histology. The diameter of the smallest follicle detected was reported using each of the three imaging methods. The number of antral follicles (antral follicle count ≥2 mm) was compared between ultrasonography and DEI.

DEI enabled the detection of 71% of follicles and 67% of CL that were detected ultrasonographically. However, DEI did not allow the detection of COCs and cell layers of the follicle wall that were visualized histologically. Luteal tissues were not easily distinguished using DEI, and DEI was inferior for differentiating follicles and CL compared with ultrasonography. The mean follicle diameter was similar between DEI (4.00 ± 0.35 mm, fresh with contrast; 9.62 ± 2.43 mm, fresh without contrast) and ultrasonography (3.85 ± 0.28 mm, fresh with contrast; 8.97 ± 2.60 mm, fresh without contrast) (P > .05). However, the mean follicle diameter was greater using both DEI (4.00 ± 0.35 mm) and ultrasonography (3.85 ± 0.28 mm) compared with histology (2.21 ± 0.38 mm; P = .01, fresh ovaries with contrast). The mean CL diameter was similar between DEI (11.64 ± 1.67 mm), ultrasonography (9.34 ± 0.35 mm), and histology (9.59 ± 0.36 mm) (P > .05). The mean diameter of the smallest follicle detected was similar between DEI (3.06 ± 0.45 mm) and ultrasonography (2.95 ± 0.74 mm); both DEI and ultrasonographic measurements were greater than histology (0.39 ± 0.04 mm, P < .0001). The mean antral follicle count was similar between ultrasonography (6.50 ± 0.71 mm, fresh with no contrast; 6.50 ± 2.50 mm, preserved with no contrast) and DEI (4.50 ± 0.50 mm, fresh with no contrast; 6.50 ± 0.50 mm, preserved with no contrast) (P > .05).

The contrast resolution of antral follicles, CL, and COCs in bovine ovaries was inferior using DEI compared with ultrasonography and histology. Alternative synchrotron techniques, such as phase-contrast computed tomography and DEI computed tomography, may prove more effective than DEI for imaging ovaries ex vivo.