Abstract
Basic and translational research in reproductive medicine can provide new insights with the application of scanning probe microscopies, such as atomic force microscopy (AFM) and scanning near-field optical microscopy (SNOM). These microscopies, which provide images with spatial resolution well beyond the optical resolution limit, enable users to achieve detailed descriptions of cell topography, inner cellular structure organization, and arrangements of single or cluster membrane proteins. A peculiar characteristic of AFM operating in force spectroscopy mode is its inherent ability to measure the interaction forces between single proteins or cells, and to quantify the mechanical properties (i.e., elasticity, viscoelasticity, and viscosity) of cells and tissues. The knowledge of the cell ultrastructure, the macromolecule organization, the protein dynamics, the investigation of biological interaction forces, and the quantification of biomechanical features can be essential clues for identifying the molecular mechanisms that govern responses in living cells. This review highlights the main findings achieved by the use of AFM and SNOM in assisted reproductive research, such as the description of gamete morphology; the quantification of mechanical properties of gametes; the role of forces in embryo development; the significance of investigating single-molecule interaction forces; the characterization of disorders of the reproductive system; and the visualization of molecular organization. New perspectives of analysis opened up by applying these techniques and the translational impacts on reproductive medicine are discussed.
Highlights
Basic research in the reproductive medicine field has led to the development of the assisted reproductive techniques (ARTs) that can overcome human infertility
Microscopy but better lateral resolution [28]. The potentialities of this technique applied to imaging of fluorescent-labeled single biomolecules are underlined by several papers [29,30], which show the ability of scanning near-field optical microscopy (SNOM) to map the molecular organization of protein clusters or lipid rafts in the cell membrane, to study their involvement in cell function
The application of electron microscopy has allowed describing in detail the ultrastructural anomalies of the human spermatozoa [34,35] and of the human oocytes, which can affect cell function [36]
Summary
During the past 30 years, scanning probe microscopies (SPM) quickly gained widespread attention, from hard materials science to biology [17]. To a first approximation: transmitted light provides information as dark field optical microscopy does in the far-field, while reflected light provides information to phase imaging, both with better spatial resolution In this way, SNOM can be exploited to acquire superresolution optical images by scanning a probe tip over a surface [27]. Microscopy but better lateral resolution [28] The potentialities of this technique applied to imaging of fluorescent-labeled single biomolecules are underlined by several papers [29,30], which show the ability of SNOM to map the molecular organization of protein clusters or lipid rafts in the cell membrane, to study their involvement in cell function. It finds better applications in the investigation of lipid membranes, cell membranes, single protein, isolated fluorophores, fibrils, DNA, and other nanosized structures [32,33]
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