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Label-free imaging and biochemical characterization of bovine sperm cells.

Ferrara MA, Di Caprio G, Managò S, De Angelis A, Sirleto L, Coppola G, De Luca AC - Biosensors (Basel) (2015)

Bottom Line: 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).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.

View Article: PubMed Central - PubMed

Affiliation: Institute for Microelectronics and Microsystems, National Research Council, Via P. Castellino, 111, 80131 Naples, Italy. antonella.ferrara@na.imm.cnr.it.

ABSTRACT
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.

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(A) Phase map; (B) Cut phase map; (C) Otsu’s method; (D) Regionprops maximum area; (E) Boundary; (F) Polygonal interpolation; (G) Filling; (H) Expansion; (I) Product phase-mask.
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biosensors-05-00141-f007: (A) Phase map; (B) Cut phase map; (C) Otsu’s method; (D) Regionprops maximum area; (E) Boundary; (F) Polygonal interpolation; (G) Filling; (H) Expansion; (I) Product phase-mask.

Mentions: With this aim, a MATLAB code was implemented, and the applied procedure is reported in Figure 7. On the zoomed phase map (Figure 7A,B), retrieved by means of the aforementioned numerical approach, the threshold is computed by applying the Otsu’s method (Figure 7C) [44]. Then, the “regionprops”, a MATLAB function, is applied to measure properties of image regions (Figure 7D), and boundary of the region of interest and its polygonal interpolation are performed (Figure 7E,F). A filling and an expansion of the selected area are done (Figure 7G,H); finally the product between the phase map (Figure 7B) and the obtained mask (Figure 7H) is implemented (Figure 7I).


Label-free imaging and biochemical characterization of bovine sperm cells.

Ferrara MA, Di Caprio G, Managò S, De Angelis A, Sirleto L, Coppola G, De Luca AC - Biosensors (Basel) (2015)

(A) Phase map; (B) Cut phase map; (C) Otsu’s method; (D) Regionprops maximum area; (E) Boundary; (F) Polygonal interpolation; (G) Filling; (H) Expansion; (I) Product phase-mask.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493542&req=5

biosensors-05-00141-f007: (A) Phase map; (B) Cut phase map; (C) Otsu’s method; (D) Regionprops maximum area; (E) Boundary; (F) Polygonal interpolation; (G) Filling; (H) Expansion; (I) Product phase-mask.
Mentions: With this aim, a MATLAB code was implemented, and the applied procedure is reported in Figure 7. On the zoomed phase map (Figure 7A,B), retrieved by means of the aforementioned numerical approach, the threshold is computed by applying the Otsu’s method (Figure 7C) [44]. Then, the “regionprops”, a MATLAB function, is applied to measure properties of image regions (Figure 7D), and boundary of the region of interest and its polygonal interpolation are performed (Figure 7E,F). A filling and an expansion of the selected area are done (Figure 7G,H); finally the product between the phase map (Figure 7B) and the obtained mask (Figure 7H) is implemented (Figure 7I).

Bottom Line: 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).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.

View Article: PubMed Central - PubMed

Affiliation: Institute for Microelectronics and Microsystems, National Research Council, Via P. Castellino, 111, 80131 Naples, Italy. antonella.ferrara@na.imm.cnr.it.

ABSTRACT
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.

Show MeSH
Related in: MedlinePlus