Digital holographic microscopy for the evaluation of human sperm structure

Abstract

The morphology of the sperm head has often been correlated with the outcome of in vitro fertilization (IVF), and has been shown to be the sole parameter in semen of value in predicting the success of intracytoplasmic sperm injection (ICSI) and intracytoplasmic morphologically selected sperm injection (IMSI). 25In this paper, we have studied whether Digital Holographic (DH) microscopy may be useful to obtain quantitative data on human sperm head structure and compared this technique to high power digitally enhanced Nomarski microscope. The main advantage of DH is that a high resolution 3-D quantitative sample imaging may be obtained thorugh numerical refocusing at different object planes without any mechanical scanning. We show that DH can furnish useful information on the 30 dimensions and structure of human spermatozoo, that cannot be revealed by conventional phase contrast microscopy. In fact, in this paper DH has been used to evaluate volume and indicate precise location of vacuoles, thus suggesting its use as an additional useful prognostic quantitative tool in assisted reproduction technology (ART).

Interferometric phase microscopy for label-free morphological evaluation of sperm cells

“…Coppola et al (10) used IPM to evaluate sperm, mainly based on the presence or absence of vacuoles. Because vacuoles form early in spermatogenesis, they may reflect defects in sperm content, and therefore, these cells should be avoided in ICSI.…”

“…This method requires a lower illumination power and presents high throughput because capturing is done in a single exposure and without scanning. Holographic imaging has been identified previously as a tool for sperm measurements (10,11). However, until recently, most IPM setups were bulky, expensive, and hard to operate.…”

Holography: The Usefulness of Digital Holographic Microscopy for Clinical Diagnostics

“…[43]. The morphology of the sperm head has often been correlated with the outcome of in vitro fertilization and has been shown to be the sole parameter in semen of value in predicting the success of intracytoplasmic sperm injection and intracytoplasmic morphologically selected sperm injection [44,45]. Indeed, DH microscopy generates useful information on the dimensions and structure of human sperm, not revealed by conventional phase-contrast microscopy, in particular the volume of vacuoles.…”

Unlabeled Semen Analysis by Means of the Holographic Imaging

“…The 3D analysis can add further information to the data provided by the traditional bi-dimensional optical techniques, allowing to better understand the relationship between the male infertility and the abnormal morphology. Due to this potentiality, the holographic approach has been also employed to analyze the human sperm characteristics [35,36]. Crha et al [35] tested about 3000 sperm cells to individuate a phase difference between spermatozoa in normo-zoospermia (NZ) and oligoasthenoteratozoospermia (OAT).…”

“…The Dale's group performed a comparison between the results of spermatozoa analyzed both by a semiautomated digitally enhanced Nomarski microscopy (DESA) and by the holographic imaging [36]. In Table 2, the values relative to five primary parameters (length, width, perimeter, area, and volume) for normal human spermatozoa are compared.…”