The application of scanning near field optical imaging to the study of human sperm morphology

Andolfi, Laura; Trevisan, Elisa; Troian, Barbara; Prato, Stefano Del; Boscolo, Rita; Giolo, Elena; Luppi, Stefania; Martinelli, Monica; Ricci, Giuseppe; Zweyer, Marina

doi:10.1186/s12951-014-0061-5

Cited by 12 publications

(8 citation statements)

References 58 publications

(51 reference statements)

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“…Although SNOM has been shown to reveal cytoplasmic structures in previous studies 12 , the SNOM-IR-FEL images obtained in this study were difficult to interpret in terms of structures. This was due to the use of a cleaved fibre for the SNOM imaging, which added significant tip artefacts to the topographical image and often off-set the topographic from the transmission images.…”

Section: Discussioncontrasting

confidence: 63%

“…SNOM has been used to investigate the localisation of molecules within cell membranes of prostate cancer cells 11 . Further research demonstrated SNOM can accurately define both the cell surface and internal structures in both healthy and anomalous sperm, including the acrosome, nucleus and the organisation of mitochondria 12 , and has demonstrated potential for single molecule imaging 13 . The application of SNOM to oesophageal cancer tissue studies provided evidence of its potentiality for cancer diagnosis 8 .…”

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confidence: 99%

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Imaging cervical cytology with scanning near-field optical microscopy (SNOM) coupled with an IR-FEL

Halliwell

Morais

Lima

et al. 2016

Sci Rep

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Cervical cancer remains a major cause of morbidity and mortality among women, especially in the developing world. Increased synthesis of proteins, lipids and nucleic acids is a pre-condition for the rapid proliferation of cancer cells. We show that scanning near-field optical microscopy, in combination with an infrared free electron laser (SNOM-IR-FEL), is able to distinguish between normal and squamous lowgrade and high-grade dyskaryosis, and between normal and mixed squamous/glandular pre-invasive and adenocarcinoma cervical lesions, at designated wavelengths associated with DNA, Amide I/II and lipids. These findings evidence the promise of the SNOM-IR-FEL technique in obtaining chemical information relevant to the detection of cervical cell abnormalities and cancer diagnosis at spatial resolutions below the diffraction limit (≥0.2 μm). We compare these results with analyses following attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy; although this latter approach has been demonstrated to detect underlying cervical atypia missed by conventional cytology, it is limited by a spatial resolution of ~3 μm to 30 μm due to the optical diffraction limit.Cervical cancer is associated with the persistent infection of high-risk types of human papillomavirus (HPV), together with other socioeconomic co-factors 1 . Screening involves cytological and histological classification of cervical cells. In the UK, cytological examination of cervical squamous cells is classified as normal, borderline or mild dyskaryosis, moderate or severe dyskaryosis and invasive cervical cancer (ICC). Histology is defined as normal, cervical intra-epithelial neoplasia (CIN): CIN1, CIN2, CIN3, or invasive cervical cancer. For atypical cells found in the glandular cells of the cervix, the pre-invasive lesion of adenocarcinoma is defined by changes termed cervical glandular intraepithelial neoplasia (CGIN), and sub-classified as low-grade cervical glandular intra-epithelial neoplasia (LGCGIN) and high-grade cervical glandular intra-epithelial neoplasia (HGCGIN). Squamous and glandular lesions may co-exist together and are defined by the level of CIN together with either LGCGIN or HGCGIN. Conventional screening is flawed as it is dependent on the subjective visual inspection of cytology; this often results in mis-diagnoses when grading samples 2 .

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Section: Discussioncontrasting

confidence: 63%

mentioning

confidence: 99%

Imaging cervical cytology with scanning near-field optical microscopy (SNOM) coupled with an IR-FEL

Halliwell

Morais

Lima

et al. 2016

Sci Rep

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“…With this technique peculiar structural features of these cells have been disclosed without staining procedures. As shown in Figure 3, the helicoidal organization of mitochondria in the midpiece region is well discernible in normal spermatozoa [94], and other structural features of healthy and anomalous spermatozoa are detected, all well resembling those observed in transmission electron microscopy images [95]. These works demonstrate the ability of SNOM to visualize inner cellular compartments with a typical contrast (transmission and reflection) for standard microscopy but with a higher resolution [94,95].…”

Section: Inner Cellular Organization Of Spermatozoasupporting

confidence: 63%

Scanning Probe Microscopies: Imaging and Biomechanics in Reproductive Medicine Research

Andolfi

Battistella

Zanetti

et al. 2021

IJMS

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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.

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“…It is seen that assisted reproduction using spermatozoa with deformities not only leads to lower fertilization and pregnancy rates, but also imposes higher risk of lesions in fetal DNA (Nikbakht & Saharkhiz, ). Recently, nanoscopic techniques are being devised to describe structural details of spermatozoa (Andolfi et al, ). Taking into account the enormous prognostic value attributed to sperm morphology in assisted reproduction, we delved into this aspect in depth.…”

Section: Discussionmentioning

confidence: 99%

Testicular germ cell apoptosis and sperm defects in mice upon long‐term high fat diet feeding

Ghosh

Mukherjee

2018

Journal Cellular Physiology

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The growing prevalence of male infertility is a matter of serious concern. One of the putative causes being nutritional excess from continuous consumption of high fat diet (HFD) leading to insulin resistance albeit the specific relationship is not fully understood. Besides, there are many contradictions in the available literature on the subject. Therefore, we sought to characterize in detail the effects of HFD upon testicular function and sperm quality in mice with particular focus on isolated testicular germ cells and spermatozoa, respectively. In this study, we treated 8-week old male Swiss albino mice with HFD for the duration of 5 months; control animals were reared on standard diet. We observed HFD induced progressive deterioration of testicular histoarchitecture leading to disruption of seminiferous tubules, increased vacuolization, and partial to complete tubular atrophy. Time dependent adverse effects on sperm count, motility, and morphology were noticed. Interestingly, numerous anomalies were detectable in sperm head and tail structures reflecting loss of reproductive capacity due to HFD. Maximal tissue and sperm damage was conspicuous at the endpoint, prompting us to examine oxidative stress markers. Enhanced intracellular reactive oxygen species (ROS) generation, augmentation of prooxidant activities, and compromised testicular antioxidant defences clearly implied conditions of oxidative stress in long-term HFD treated mice. This was concomitant with the onset of abnormally enhanced testicular germ cell apoptosis involving the mitochondrial intrinsic pathway. Thus, our findings revealed that ROS mediated deregulation of testicular germ cell apoptosis is critical in male reproductive impairment due to diet induced obesity.

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The application of scanning near field optical imaging to the study of human sperm morphology

Cited by 12 publications

References 58 publications

Imaging cervical cytology with scanning near-field optical microscopy (SNOM) coupled with an IR-FEL

Imaging cervical cytology with scanning near-field optical microscopy (SNOM) coupled with an IR-FEL

Scanning Probe Microscopies: Imaging and Biomechanics in Reproductive Medicine Research

Testicular germ cell apoptosis and sperm defects in mice upon long‐term high fat diet feeding

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