Limits...
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.

Show MeSH

Related in: MedlinePlus

(A) Acquired hologram, a region is enhanced in order to show the interference pattern (inset); (B) Pseudo 3D representation of the phase map of a bovine spermatozoon obtained by digital holography microscopy.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4493542&req=5

biosensors-05-00141-f003: (A) Acquired hologram, a region is enhanced in order to show the interference pattern (inset); (B) Pseudo 3D representation of the phase map of a bovine spermatozoon obtained by digital holography microscopy.

Mentions: The morphology of bovine sperm cells and the integrity of their structures were investigated by DH microscopy. An example of an acquired hologram is reported in Figure 3A, where the inset shows the intensity of the fringe pattern due to the superimposition of the object and reference beams. The reconstructed three-dimensional shape of a bovine spermatozoon is reported as a pseudocolor plot in Figure 3B. Quantitative information about the thickness of the sample under test can be obtained by Equation (2), by the phase-contrast map of the sample and considering a uniform refractive index of about 1.35, as reported in literature [33].


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) Acquired hologram, a region is enhanced in order to show the interference pattern (inset); (B) Pseudo 3D representation of the phase map of a bovine spermatozoon obtained by digital holography microscopy.
© Copyright Policy
Related In: Results  -  Collection

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

biosensors-05-00141-f003: (A) Acquired hologram, a region is enhanced in order to show the interference pattern (inset); (B) Pseudo 3D representation of the phase map of a bovine spermatozoon obtained by digital holography microscopy.
Mentions: The morphology of bovine sperm cells and the integrity of their structures were investigated by DH microscopy. An example of an acquired hologram is reported in Figure 3A, where the inset shows the intensity of the fringe pattern due to the superimposition of the object and reference beams. The reconstructed three-dimensional shape of a bovine spermatozoon is reported as a pseudocolor plot in Figure 3B. Quantitative information about the thickness of the sample under test can be obtained by Equation (2), by the phase-contrast map of the sample and considering a uniform refractive index of about 1.35, as reported in literature [33].

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