G. Coppola

A digital holographic microscope for complete characterization of microelectromechanical systems

Digital holographic microscopy (DHM) can be described as a non-invasive metrological tool for inspection and characterization of microelectromechanical structures (MEMS). DHM is a quick, non-contact and non-invasive technique that can offer a high resolution in both lateral and vertical directions. It has been employed for the characterization of the undesired out-of-plane deformations due to the residual stresses introduced by technological processes. The characterization of these deformations is helpful in studying and understanding the effect of residual stress on the deformation of a single microstructure. To that end, MEMS with different geometries and shapes, such as cantilever beams, bridges and membranes, have been characterized. Moreover, DHM has been applied efficiently to evaluate variations of the structure profile due to some external effects. As an example, the characterization of a cantilever subjected to a thermal process has been described. The results reported show that DHM is a useful non-invasive method for characterizing and developing reliable MEMS.

Identification of bovine sperm head for morphometry analysis in quantitative phase-contrast holographic microscopy

An investigation is reported of the identification and measurement of region of interest (ROI) in quantitative phase-contrast maps of biological cells by digital holographic microscopy. In particular, two different methods have been developed for in vitro bull sperm head morphometry analysis. We show that semen analysis can be accomplished by means of the proposed techniques . Extraction and measurement of various parameters are performed. It is demonstrated that both proposed methods are efficient to skim the data set in a preselective analysis for discarding anomalous data.

Digital self-referencing quantitative phase microscopy by wavefront folding in holographic image reconstruction.

A completely numerical method, named digital self-referencing holography, is described to easily accomplish a quantitative phase microscopy for microfluidic devices by a digital holographic microscope. The approach works through an appropriate numerical manipulation of the retrieved complex wavefront. The self-referencing is obtained by folding the retrieved wavefront in the image plane. The folding operation allows us to obtain the correct phase map by subtracting from the complex region of interest a flat area outside the microfluidic channel. To demonstrate the effectiveness of the method, quantitative phase maps of bovine spermatozoa and in vitro cells are retrieved.

4D tracking of clinical seminal samples for quantitative characterization of motility parameters

In this paper we investigate the use of a digital holographic microscope, with partial spatial coherent illumination, for the automated detection and tracking of spermatozoa. This in vitro technique for the analysis of quantitative parameters is useful for assessment of semen quality. In fact, thanks to the capabilities of digital holography, the developed algorithm allows us to resolve in-focus amplitude and phase maps of the cells under study, independently of focal plane of the sample image. We have characterized cell motility on clinical samples of seminal fluid. In particular, anomalous sperm cells were characterized and the quantitative motility parameters were compared to those of normal sperm.

Quantitative Label-Free Animal Sperm Imaging by Means of Digital Holographic Microscopy

A digital holographic microscope (DHM) has been employed in the retrieval and analysis of morphological images of bovines sperm cells. Digital holography is a noncontact technique capable of investigating the shape of the sample without altering its characteristics and has been used for the first time in retrieving quantitative morphological information of sperm cells. Different spermatozoa have been analyzed by means of this technique allowing us to obtain 3-D images with precise topographical details and valuable information about morphological defects, provided with biological considerations. Moreover, by making use of a microfluidic system, the digital holographic technique has been employed to analyze unstained spermatozoa in their natural physiological surroundings. Detailed information on morphological images of spermatozoa acquired by DHM is expected to provide a better understanding of various reproductive pathways, which, in turn, can help in improving infertility management. This could constitute the basis of an alternative method for the zoothecnic industry aimed at the investigation of morphological features and the sorting of the motile sperm cells.

Surface plasmon resonance optical cavity enhanced refractive index sensing

We report on a method for surface plasmon resonance (SPR) refractive index sensing based on direct time-domain measurements. An optical resonator is built around an SPR sensor, and its photon lifetime is measured as a function of loss induced by refractive index variations. The method does not rely on any spectroscopic analysis or direct intensity measurement. Time-domain measurements are practically immune to light intensity fluctuations and thus lead to high resolution. A proof of concept experiment is carried out in which a sensor response to liquid samples of different refractive indices is measured. A refractive index resolution of the current system, extrapolated from the reproducibility of cavity-decay time determinations over 133 s, is found to be about 10(-5) RIU. The possibility of long-term averaging suggests that measurements with a resolution better than 10(-7) RIU/√Hz are within reach.

Asymmetric MSM sub-bandgap all-silicon photodetector with low dark current

Design, fabrication, and characterization of an asymmetric metal-semiconductor-metal photodetector, based on internal photoemission effect and integrated into a silicon-on-insulator waveguide, are reported. For this photodetector, a responsivity of 4.5 mA/W has been measured at 1550 nm, making it suitable for power monitoring applications. Because the absorbing metal is deposited strictly around the vertical output facet of the waveguide, a very small contact area of about 3 µm2 is obtained and a transit-time-limited bandwidth of about 1 GHz is demonstrated. Taking advantage of this small area and electrode asymmetry, a significant reduction in the dark current (2.2 nA at -21 V) is achieved. Interestingly, applying reverse voltage, the photodetector is able to tune its cut-off wavelength, extending its range of application into the MID infrared regime.

Phase map retrieval in digital holography: avoiding the undersampling effect by a lateral shear approach

In digital holography (DH) the numerical reconstruction of the whole wavefront allows one to extract the wrapped phase map mod, 2 pi. It can occur that the reconstructed wrapped phase map in the image plane is undersampled because of the limited pixel size in that plane. In such a case the phase distribution cannot be retrieved correctly by the usual unwrapping procedures. We show that the use of the digital lateral-shearing interferometry approach in DH provides the correct reconstruction of the phase map in the image plane, even in extreme cases where the phase profile changes very rapidly. We demonstrate the effectiveness of the method in a particular case where the profile of a highly curved silicon microelectromechanical system membrane has to be reconstructed.

Surface topography of microstructures in lithium niobate by digital holographic microscopy

We report here on the application of digital holographic microscopy as a metrological tool for the inspection and the micro-topography reconstruction of different microstructures fabricated in bulk lithium niobate by differential etching of reversed ferroelectric domain patterned crystals. These structures have a range of applications in optical ridge waveguides, alignment structures, V-grooves, micro-tips and micro-cantilever beams and precise control of the surface quality and topography is required. The technique allows us to obtain digitally a high-fidelity surface topography description of the specimen with only one image acquisition allowing us to have relatively simple and compact set-ups able to give quantitative information on object morphology. The advantages of this technique compared to traditional microscopy are discussed.

Non-invasive sex assessment in bovine semen by Raman spectroscopy

X- and Y-chromosome-bearing sperm cell sorting is of great interest, especially for animal production management systems and genetic improvement programs. Here, we demonstrate an optical method based on Raman spectroscopy to separate X- and Y-chromosome-bearing sperm cells, overcoming many of the limitations associated with current sex-sorting protocols. A priori Raman imaging of bull spermatozoa was utilized to select the sampling points (head-neck region), which were then used to discriminate cells based on a spectral classification model. Main variations of Raman peaks associated with the DNA content were observed together with a variation due to the sex membrane proteins. Next, we used principal component analysis to determine the efficiency of our device as a cell sorting method. The results (>90% accuracy) demonstrated that Raman spectroscopy is a powerful candidate for the development of a highly efficient, non-invasive, and non-destructive tool for sperm sexing.