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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) Profile of the head along the line DD' illustrated in (B).
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biosensors-05-00141-f004: (A) Profile of the head along the line DD' illustrated in (B).

Mentions: Figure 3B shows an interesting feature, common to many analyzed spermatozoa: a “protuberance” on the post-acrosomal region of the head. This alteration is well visible in the profile reported in Figure 4A relative to the line DD' shown in Figure 4B and it has been highlighted for the first time in our previous paper [12]. Among possible explanations of the “protuberance” nature, an artifact due to a possible different refractive index between the post-acrosomal region, (containing almost exclusively highly compact chromatin) and the acrosome should be excluded. Thus, the same characteristic has been observed in images acquired by confocal dual core confocal microscope (DCM) 3-D Leica microscope and reported in our previous paper [12]. However, both holographic and confocal images only reveal the presence of morphological defect, while they are not able to identify its biochemical constitution. A first explanation for the estimated area and volume variation is that the “protuberance” highlighted by DH microscopy could be due to the presence of the centrioles into the structures connecting the tail to the head.


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) Profile of the head along the line DD' illustrated in (B).
© Copyright Policy
Related In: Results  -  Collection

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

biosensors-05-00141-f004: (A) Profile of the head along the line DD' illustrated in (B).
Mentions: Figure 3B shows an interesting feature, common to many analyzed spermatozoa: a “protuberance” on the post-acrosomal region of the head. This alteration is well visible in the profile reported in Figure 4A relative to the line DD' shown in Figure 4B and it has been highlighted for the first time in our previous paper [12]. Among possible explanations of the “protuberance” nature, an artifact due to a possible different refractive index between the post-acrosomal region, (containing almost exclusively highly compact chromatin) and the acrosome should be excluded. Thus, the same characteristic has been observed in images acquired by confocal dual core confocal microscope (DCM) 3-D Leica microscope and reported in our previous paper [12]. However, both holographic and confocal images only reveal the presence of morphological defect, while they are not able to identify its biochemical constitution. A first explanation for the estimated area and volume variation is that the “protuberance” highlighted by DH microscopy could be due to the presence of the centrioles into the structures connecting the tail to the head.

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