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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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Related in: MedlinePlus

(A) Histogram of the measured head volume for both 500 X- and 500 Y-bearing sperm cells and the correspondent fit; (B) Mean value of the head volume obtained by the fit.
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biosensors-05-00141-f008: (A) Histogram of the measured head volume for both 500 X- and 500 Y-bearing sperm cells and the correspondent fit; (B) Mean value of the head volume obtained by the fit.

Mentions: Measures of the volume of sperm head are performed on 500 X- and 500 Y-bearing sperm cells from two different bulls to take into accounts the variation from cell to cell and bull to bull. Results are reported in Figure 8A and summarized in Figure 8B. It is evident that, even if the mean value of the volume is bigger for X-bearing sperm cells respect to Y-bearing sperm cells, as expected, the X- and Y-Gaussian are widely overlapped. Indeed, considering that the standard deviation in volume differences (2.5%) is comparable with the differences in DNA content (3.8% in bull sperm), these parameters do not allow the unequivocal identification of the two cell populations. The label-free identification of DNA content, plasma-membrane proteins or biochemical signatures for either X- or Y-bearing sperm cells would offer great opportunities for separation.


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) Histogram of the measured head volume for both 500 X- and 500 Y-bearing sperm cells and the correspondent fit; (B) Mean value of the head volume obtained by the fit.
© Copyright Policy
Related In: Results  -  Collection

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

biosensors-05-00141-f008: (A) Histogram of the measured head volume for both 500 X- and 500 Y-bearing sperm cells and the correspondent fit; (B) Mean value of the head volume obtained by the fit.
Mentions: Measures of the volume of sperm head are performed on 500 X- and 500 Y-bearing sperm cells from two different bulls to take into accounts the variation from cell to cell and bull to bull. Results are reported in Figure 8A and summarized in Figure 8B. It is evident that, even if the mean value of the volume is bigger for X-bearing sperm cells respect to Y-bearing sperm cells, as expected, the X- and Y-Gaussian are widely overlapped. Indeed, considering that the standard deviation in volume differences (2.5%) is comparable with the differences in DNA content (3.8% in bull sperm), these parameters do not allow the unequivocal identification of the two cell populations. The label-free identification of DNA content, plasma-membrane proteins or biochemical signatures for either X- or Y-bearing sperm cells would offer great opportunities for separation.

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