Advancing ovarian folliculometry with selective plane illumination microscopy.

Hsiao-Chun Amy Lin,Angelika Schnieke,Daniel Razansky,Rahul Dutta,Subhamoy Mandal,Alexander Kind

doi:10.1038/srep38057

Hsiao-Chun Amy Lin, Angelika Schnieke + Show 4 more

Open Access

https://doi.org/10.1038/srep38057

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Abstract

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. The large field of view and fast acquisition speed of our SPIM system enables rendering of volumetric image stacks from intact whole porcine ovarian follicles, clearly visualizing follicular features including follicle volume and average diameter (70 μm–2.5 mm), their spherical asymmetry parameters, size of developing cumulus oophorus complexes (40 μm–110 μm), and follicular wall thickness (90 μm–120 μm). Follicles at all developmental stages were identified. A distribution of the theca thickness was measured for each follicle, and a relationship between these distributions and the stages of follicular development was discerned. 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.

Highlights

The incidence of ovarian ailments such as premature ovarian failure (POF) and polycystic ovary syndrome (PCOS) has risen in recent years, constituting a major cause of female subfertility in the modern world[1,2]
The study we describe explore the use of selective plane illumination microscopy (SPIM) to gain accurate and informative folliculometric data from scanned ovaries
An SPIM system was built in-house to optimally support the imaging of ovarian follicles

Summary

Introduction

The incidence of ovarian ailments such as premature ovarian failure (POF) and polycystic ovary syndrome (PCOS) has risen in recent years, constituting a major cause of female subfertility in the modern world[1,2]. The diameter of the ovarian follicle and the developing oocyte within, and the thickness of the follicular wall are important parameters for clinical management of ovarian ailments. The study we describe explore the use of selective plane illumination microscopy (SPIM) to gain accurate and informative folliculometric data from scanned ovaries. SPIM is time- and labour-efficient and well suited for producing high resolution imaging stacks of 3D biological samples. It has been used for in vivo studies of naturally transparent biological specimens, such as zebrafish embryos and Drosophila pupae[9,10], as well as samples rendered optically transparent by chemical clearing[11,12]. Pig was chosen as the model organism for this study because of its similarity to humans in terms of genetics, anatomy, and physiology

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