Non-invasive sex assessment in bovine semen by Raman spectroscopy

Luca, Anna Chiara De; Managò, Stefano; Ferrara, Maria Antonietta; Rendina, Ivo; Sirleto, L.; Puglisi, Roberto; Balduzzi, Donatella; Galli, Andrea; Ferraro, Pietro; Coppola, Giuseppe

doi:10.1088/1612-2011/11/5/055604

Cited by 43 publications

(71 citation statements)

References 45 publications

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“…To measure the discrimination ability of the proposed approach, we build up the confusion matrix, using the leave-one-out classification method [20], and the classification results are shown in Table 2. By analysing the diagonal values of the confusion matrix for X and Y bovine sperm cells, we obtain an accuracy of 90.6% for differentiating between bovine X-bearing or V-bearing sperm , which is comparable to the accuracy (90-94%) achievable with FACS [16]. …”

Section: B Pea Analysismentioning

confidence: 66%

“…EXPERIMENT AL Spectra were acquired using a home-built Raman microscope ( Fig.l) [16]. A 532 nm diode laser (Opus, Laser Quantum, Maximum Power 2W) is used for the sample excitation.…”

Section: IImentioning

confidence: 99%

“…The bovine sperm cells were prepared by the Institute "Lazzaro Spallanzani" after fixation in suspension of the seminal material with 0.2% glutaraldehyde solution in phosphate buffered saline (PBS) without calcium and magnesium (1:3 v/v) [16,17]. A drop with volume 4 ilL has been deposed on a quartz microscope slide (20 mm x 40 mm, 1 mm thickness), and then, covered with a quartz coverslip (20 mm x 20 mm, 150 11m thickness).…”

Section: B Sample Preparationmentioning

confidence: 99%

“…We demonstrate that a spectroscopic signature, combination of the spectral component in the sperm (DNA, proteins, lipid etc. ), can be used to identify the biochemical components and differences between X-and Y-sperm cells [16].…”

Section: Introductionmentioning

confidence: 99%

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Raman sex sorting of bovine spermatozoa

Managò

Coppola

Ferrara

et al. 2014

2014 Fotonica AEIT Italian Conference on Photonics Technologies

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This work relates to the problem of sex predetermination in animals based on the separation of X-and Y -bearing sperm cells before insemination. We developed a device for efficient, non-destructive and label-free sorting of X and Y -bearing sperm cells based on their Raman spectroscopic characteristics. Raman spectroscopy (RS) is a non-invasive technique that allows the biochemical analysis of the cellular components and the characterization of molecular structure from their stretching and bending vibrational transitions. RS offers detailed information on the conformation, composition and molecular interactions of important cellular macromolecules, such as DNA, proteins and lipids, and can be used to characterize and study individual living cells.In this paper, we present a Raman spectroscopy-based method for selective and sensitive discrimination of X-and Y bearing bovine sperm cells.

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Section: B Pea Analysismentioning

confidence: 66%

“…EXPERIMENT AL Spectra were acquired using a home-built Raman microscope ( Fig.l) [16]. A 532 nm diode laser (Opus, Laser Quantum, Maximum Power 2W) is used for the sample excitation.…”

Section: IImentioning

confidence: 99%

Section: B Sample Preparationmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Raman sex sorting of bovine spermatozoa

Managò

Coppola

Ferrara

et al. 2014

2014 Fotonica AEIT Italian Conference on Photonics Technologies

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“…This paper demonstrates the first experimental application of this approach to the case of imaging spontaneous Raman scattering. In particular, Raman scattering is a vibrational microscopy method offering a label-free imaging contrast mechanism capable of probing non-invasively the chemical composition of biological and inert materials at microscopic scales [8][9][10][11][12]. It has promise for many applications yet has been hampered by long acquisition times which in turn has resulted in researchers resorting typically to more complex nonlinear vibrational spectroscopy methods.…”

Section: Introductionmentioning

confidence: 99%

Nonredundant Raman imaging using optical eigenmodes

Kosmeier¹,

Zolotovskaya²,

Luca³

et al. 2014

Optica

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Various forms of imaging schemes have emerged over the last decade that are based on correlating variations in incident illuminating light fields to the outputs of single 'bucket' detectors. However, to date, the role of the orthogonality of the illumination fields has largely been overlooked and furthermore, the field has not progressed beyond bright field imaging. By exploiting the concept of orthogonal illuminating fields, we demonstrate the application of optical eigenmodes (OEi) to wide field, scan-free spontaneous Raman imaging, which is notoriously slow in wide-field mode. The OEi approach enables a form of indirect imaging that exploits both phase and amplitude in image reconstruction. The use of orthogonality enables us to non-redundantly illuminate the sample and, in particular, use a subset of illuminating modes to obtain the majority of information from the sample, thus minimising any photobleaching or damage of the sample. The crucial incorporation of phase, in addition to amplitude, in the imaging process significantly reduces background noise and results in an improved SNR for the image while reducing the number of illuminations. As an example we can reconstruct images of a surface-enhanced Raman spectroscopy (SERS) active sample with approximately an order of magnitude fewer acquisitions. This generic approach may readily be applied to other imaging modalities such as fluorescence microscopy or nonlinear vibrational microscopy.

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Raman detection and identification of normal and leukemic hematopoietic cells

Managò

Mirabelli

Napolitano

2018

Journal of Biophotonics

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The analysis of leukocytes of peripheral blood is a crucial step in hematologic exams commonly used for disease diagnosis and, typically, requires molecular labelling. In addition, only a detailed, laborious phenotypic analysis allows identifying the presence and stage of specific pathologies such as leukemia. Most of the biochemical information is lost in the routine blood tests. In the present study, we tackle 2 important issues of label-free biochemical identification and classification of leukocytes using Raman spectroscopy (RS). First, we demonstrate that leukocyte subpopulations of lymphocytes (B, T and NK cells), monocytes and granulocytes can be identified by the unsupervised statistical approach of principal component analysis and classified by linear discriminant analysis with approximately 99% of accuracy. Second, we apply the same procedure to identify and discriminate normal B cells and transformed MN60 lymphocyte leukemic cell lines. In addition, we demonstrate that RS can be efficiently used for monitoring the cell response to low-dose chemotherapy treatment, experimentally eliciting the sensitivity to a dose-dependent cell response, which is of fundamental importance to determine the efficacy of any treatment. These results largely expand established Raman-based research protocols for label-free analysis of white blood cells, leukemic cells and chemotherapy treatment follow-up.

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Non-invasive sex assessment in bovine semen by Raman spectroscopy

Cited by 43 publications

References 45 publications

Raman sex sorting of bovine spermatozoa

Raman sex sorting of bovine spermatozoa

Nonredundant Raman imaging using optical eigenmodes

Raman detection and identification of normal and leukemic hematopoietic cells

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