Determination of ovarian status and follicle monitoring are common methods of diagnosing female infertility. We
evaluated the suitability of selective plane illumination microscopy (SPIM) for the study of ovarian follicles. Owing to
the large field of view and fast acquisition speed of our newly developed SPIM system, volumetric image stacks from
entire intact samples of pig ovaries have been rendered demonstrating clearly discernible follicular features like follicle
diameters (70 μm - 2.5 mm), size of developing Cumulus oophorus complexes (COC ) (40 μm - 110 μm), and follicular
wall thicknesses (90 μm-120 μm). The observation of clearly distinguishable COCs protruding into the follicular antrum
was also shown possible, and correlation with the developmental stage of the follicles was determined. Follicles of all
developmental stages were identified, and even the small primordial follicle clusters forming the egg nest could be
observed. The ability of the system to non-destructively generate sub-cellular resolution 3D images of developing
follicles, with excellent image contrast and high throughput capacity compared to conventional histology, suggests that it
can be used to monitor follicular development and identify structural abnormalities indicative of ovarian ailments.
Accurate folliculometric measurements provided by SPIM images can immensely help the understanding of ovarian
physiology and provide important information for the proper management of ovarian diseases.
|