Label-free imaging and biochemical characterization of bovine sperm cells

Ferrara, Maria Antonietta and Di Caprio, Giuseppe and Managò, Stefano and De Angelis, Annalisa and Sirleto, Luigi and Coppola, Giuseppe and De Luca, Anna Chiara (2015) Label-free imaging and biochemical characterization of bovine sperm cells. Biosensors, 5 (2). pp. 141-157. ISSN 2079-6374 (

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A full label-free morphological and biochemical characterization is desirable to select spermatozoa during preparation for artificial insemination. In order to study these fundamental parameters, we take advantage of two attractive techniques: digital holography (DH) and Raman spectroscopy (RS). DH presents new opportunities for studying morphological aspect of cells and tissues non-invasively, quantitatively and without the need for staining or tagging, while RS is a very specific technique allowing the biochemical analysis of cellular components with a spatial resolution in the sub-micrometer range. In this paper, morphological and biochemical bovine sperm cell alterations were studied using these techniques. In addition, a complementary DH and RS study was performed to identify X- and Y-chromosome-bearing sperm cells. We demonstrate that the two techniques together are a powerful and highly efficient tool elucidating some important criterions for sperm morphological selection and sex-identification, overcoming many of the limitations associated with existing protocols.