Label-Free Imaging and Biochemical Characterization of Bovine Sperm Cells

Ferrara, Maria Antonietta; Caprio, Giuseppe Di; Managò, Stefano; Angelis, Annalisa De; Sirleto, L.; Coppola, Giuseppe

doi:10.3390/bios5020141

Cited by 41 publications

(62 citation statements)

References 48 publications

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“…A scheme of the DH system used in this work is shown in our previous paper 8,26 . Briefly, the light source was a 633 nm He-Ne laser.…”

Section: Experimental Set Upmentioning

confidence: 99%

“…DH microscopy approach was used to investigate the morphology and structure integrity of the bull spermatozoa 26 . An example of an acquired hologram is reported in figure 2(a), where the inset shows the intensity of the fringe pattern due to the superimposition of the object and reference beams.…”

Section: Morphological Characterization Of Bovine Sperm Cellsmentioning

confidence: 99%

“…When the morphological analysis needs to be integrated with biochemical information, RS can be efficiently used. Indeed, RS is a sensitive and non-invasive method with resolution in the sub-micrometer range that allows the chemical assessment of single cells by detecting their biomolecule vibrations, serving as a cellular intrinsic 'fingerprint' 3,[15][16][17][18][19][20][21][22][23][24][25][26] . It can be also used to provide pseudo-color images according to the Raman spectral band intensities allowing the identification of cell phenotype and physiological state [23][24][25][26][27][28][29][30][31][32] .…”

Section: Introductionmentioning

confidence: 99%

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Spermatozoa quality assessment: a combined holographic and Raman microscopy approach

Angelis¹,

Ferrara²,

Caprio

et al. 2015

SPIE Proceedings

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Semen analysis is widely used as diagnostic tool for assessing male fertility, controlling and managing the animal reproduction. The most important parameters measured in a semen analysis are the morphology and biochemical alterations. For obtaining such information, non-invasive, label-free and non-destructive techniques have to be used. Digital Holography (DH) combined with Raman Spectroscopy (RS) could represent the perfect candidate for a rapid, non-destructive and high-sensitive morphological and biochemical sperm cell analysis. In this study, DH-RS combined approach is used for a complete analysis of single bovine spermatozoa. High-resolution images of bovine sperm have been obtained by DH microscopy from the reconstruction of a single acquired hologram, highlighting in some cases morphological alterations. Quantitative 3D reconstructions of sperm head, both normal and anomalous, have been studied and an unexpected structure of the post-acrosomal region of the head has been detected. Such anomalies have been also confirmed by Raman imaging analysis, suggesting the protein vibrations as associated Raman marker of the defect.

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“…A scheme of the DH system used in this work is shown in our previous paper 8,26 . Briefly, the light source was a 633 nm He-Ne laser.…”

Section: Experimental Set Upmentioning

confidence: 99%

Section: Morphological Characterization Of Bovine Sperm Cellsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Spermatozoa quality assessment: a combined holographic and Raman microscopy approach

Angelis¹,

Ferrara²,

Caprio

et al. 2015

SPIE Proceedings

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“…Both of the techniques were synchronised and operated simultaneously. An analysis system combining laser Raman spectroscopy and holography was previously used by Ferrara et al (2015Ferrara et al ( , 2016 as well as by Pavillon and Smith (2015) to study biological samples, e.g. the morphology and molecular composition of cells and cell components.…”

Section: Introductionmentioning

confidence: 99%

Simultaneous in situ characterisation of bubble dynamics and a spatially resolved concentration profile: a combined Mach–Zehnder holography and confocal Raman-spectroscopy sensor system

Guhathakurta

Schurr

Rinke

et al. 2017

J. Sens. Sens. Syst.

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Abstract. For a reaction between a gaseous phase and a liquid phase, the interaction between the hydrodynamic conditions, mass transport and reaction kinetics plays a crucial role with respect to the conversion and selectivity of the process. Within this work, a sensor system was developed to simultaneously characterise the bubble dynamics and the localised concentration measurement around the bubbles. The sensor system is a combination of a digital Mach-Zehnder holography subsystem to measure bubble dynamics and a confocal Raman-spectroscopy subsystem to measure localised concentration. The combined system was used to investigate the chemical absorption of CO 2 bubbles in caustic soda in microchannels. The proposed set-up is explained and characterised in detail and the experimental results are presented, illustrating the capability of the sensor system to simultaneously measure the localised concentration of the carbonate ion with a good limit of detection and the 3-D position of the bubble with respect to the spot where the concentration was measured.

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“…QPI is an emerging method that provides quantitative information about transparent specimens, with nanoscale sensitivity and in label-free mode 9 . Various implementations of QPI have been used recently to study the topography of gametes, assuming a certain refractive index [10][11][12][13][14][15][16] . Here, for the first-time to our knowledge, we measure independently the thickness and refractive index of the cells.…”

mentioning

confidence: 99%

Topography and refractometry of sperm cells using SLIM

Liu

Kandel

Rubessa

et al. 2017

Preprint

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Abstract. Characterization of spermatozoon viability is a common test in treating infertility. Recently, it has been shown that label-free, phase-sensitive imaging can provide a valuable alternative for this type of assay. Here, we employ spatial light interference microscopy (SLIM) to decouple the thickness and refractive index information of individual cells. This procedure was enabled by quantitative phase imaging cells on media of two different refractive indices and using a numerical tool to remove the curvature from the cell tails. This way, we achieved ensemble averaging of topography and refractometry of 100 cells in each of the two groups. The results show that the thickness profile of the cell tail goes down to 150 nm and the refractive index can reach values of 1.6 close to the head.

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Label-Free Imaging and Biochemical Characterization of Bovine Sperm Cells

Cited by 41 publications

References 48 publications

Spermatozoa quality assessment: a combined holographic and Raman microscopy approach

Spermatozoa quality assessment: a combined holographic and Raman microscopy approach

Simultaneous in situ characterisation of bubble dynamics and a spatially resolved concentration profile: a combined Mach–Zehnder holography and confocal Raman-spectroscopy sensor system

Topography and refractometry of sperm cells using SLIM

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