Lisa Miccio

Direct full compensation of the aberrations in quantitative phase microscopy of thin objects by a single digital hologram

Aberrations and the distortions due to the imaging optics can be compensated in quantitative phase microscopy of thin phase objects by digital holography using a single hologram. The reconstructed quantitative phase microscopy phase distribution map can be directly corrected in the reconstructed image plane by a numerical method. To remove this unwanted aberration, in the special case of thin objects, the authors perform a two-dimensional fit with the Zernike polynomials of the reconstructed unwrapped phase. Subtraction of the fitted polynomial from the original phase map gives quantitative phase microscopy phase map free of aberrations.

Liquid micro-lens array activated by selective electrowetting on lithium niobate substrates.

Lens effect was obtained in an open microfluidic system by using a thin layer of liquid on a polar electric crystal like LiNbO3. An array of liquid micro-lenses was generated by electrowetting effect in pyroelectric periodically poled crystals. Compared to conventional electrowetting devices, the pyroelectric effect allowed to have an electrode-less and circuit-less configuration. An interferometric technique was used to characterize the curvature of the micro-lenses and the corresponding results are presented and discussed. The preliminary results concerning the imaging capability of the micro-lens array are also reported.

Full Color 3-D Imaging by Digital Holography and Removal of Chromatic Aberrations

In color digital holography the recording and reconstruction of three holograms, by an optical setup operating in multi-wavelength mode, is necessary. Incorrect superposition of different images at various wavelengths may occur due to the presence of chromatic aberrations. We demonstrate that the compensation of the chromatic aberrations can be achieved by using all of the information contained in the multi-wavelength digital holograms. Chromatic aberration is extracted from phase maps reconstructed at different wavelengths, resulting in perfect full color amplitude reconstructions. The method has important applications in color 3-D holographic imaging and display.

Tunable liquid microlens arrays in electrode-less configuration and their accurate characterization by interference microscopy

A special class of tunable liquid microlenses is presented here. The microlenses are generated by an electrowetting effect under an electrode-less configuration and they exhibit two different regimes that are named here as separated lens regime (SLR) and wave-like lens regime (WLR). The lens effect is induced by the pyroelectricity of polar dielectric crystals, as was proved in principle in a previous work by the same authors (S. Grilli et al., Opt. Express 16, 8084, 2008). Compared to that work, the improvements to the experimental set-up and procedure allow to reveal the two lens regimes which exhibit different optical properties. A digital holography technique is used to reconstruct the transmitted wavefront during focusing and a focal length variation in the millimetre range is observed. The tunability of such microlenses could be of great interest to the field of micro-optics thanks to the possibility to achieve focus tuning without moving parts and thus favouring the miniaturization of the optical systems.

Recent advances in holographic 3D particle tracking

Particle tracking is a fundamental technique for investigating a variety of biophysical processes, from intracellular dynamics to the characterization of cell motility and migration. However, observing three-dimensional (3D) trajectories of particles is in general a challenging task in classical microscopy owing to the limited imaging depth of field of commercial optical microscopes, which represents a serious drawback for the analysis of time-lapse microscopy image data. Therefore, numerous automated particle-tracking approaches have been developed by many research groups around the world. Recently, digital holography (DH) in microscopy has rapidly gained credit as one of the elective techniques for these applications, mainly due to the uniqueness of the DH to provide a posteriori quantitative multiple refocusing capability and phase-contrast imaging. Starting from this paradigm, a huge amount of 3D holographic tracking approaches have been conceived and investigated for applications in various branches of science, including optofluids, microfluidics, biomedical microscopy, cell mechano-trasduction, and cell migration. Since a wider community of readers could be interested in such a review, i.e., not only scientists working in the fields of optics and photonics but also users of particle-tracking tools, it should be very beneficial to provide a complete review of state-of-the-art holographic 3D particle-tracking methods and their applications in bio-microfluidics.

Imaging live humans through smoke and flames using far-infrared digital holography

The ability to see behind flames is a key challenge for the industrial field and particularly for the safety field. Development of new technologies to detect live people through smoke and flames in fire scenes is an extremely desirable goal since it can save human lives. The latest technologies, including equipment adopted by fire departments, use infrared bolometers for infrared digital cameras that allow users to see through smoke. However, such detectors are blinded by flame-emitted radiation. Here we show a completely different approach that makes use of lensless digital holography technology in the infrared range for successful imaging through smoke and flames. Notably, we demonstrate that digital holography with a cw laser allows the recording of dynamic human-size targets. In this work, easy detection of live, moving people is achieved through both smoke and flames, thus demonstrating the capability of digital holography at 10.6 μm.

3D morphometry of red blood cells by digital holography

Three dimensional (3D) morphometric analysis of flowing and not‐adherent cells is an important aspect for diagnostic purposes. However, diagnostics tools need to be quantitative, label‐free and, as much as possible, accurate. Recently, a simple holographic approach, based on shape from silhouette algorithm, has been demonstrated for accurate calculation of cells biovolume and displaying their 3D shapes. Such approach has been adopted in combination with holographic optical tweezers and successfully applied to cells with convex shape. Nevertheless, unfortunately, the method fails in case of specimen with concave surfaces. Here, we propose an effective approach to achieve correct 3D shape measurement that can be extended in case of cells having concave surfaces, thus overcoming the limit of the previous technique. We prove the new procedure for healthy red blood cells (RBCs) (i.e., discocytes) having a concave surface in their central region. Comparative analysis of experimental results with a theoretical 3D geometrical model of RBC is discussed in order to evaluate accuracy of the proposed approach. Finally, we show that the method can be also useful to classify, in terms of morphology, different varieties of RBCs.

Tomographic flow cytometry by digital holography

High-throughput single-cell analysis is a challenging task. Label-free tomographic phase microscopy is an excellent candidate to perform this task. However, in-line tomography is very difficult to implement in practice because it requires a complex set-up for rotating the sample and examining the cell along several directions. We demonstrate that by exploiting the random rolling of cells while they are flowing along a microfluidic channel, it is possible to obtain in-line phase-contrast tomography, if smart strategies for wavefront analysis are adopted. In fact, surprisingly, a priori knowledge of the three-dimensional position and orientation of rotating cells is no longer needed because this information can be completely retrieved through digital holography wavefront numerical analysis. This approach makes continuous-flow cytotomography suitable for practical operation in real-world, single-cell analysis and with a substantial simplification of the optical system; that is, no mechanical scanning or multi-direction probing is required. A demonstration is given for two completely different classes of biosamples: red blood cells and diatom algae. An accurate characterization of both types of cells is reported, despite their very different nature and material content, thus showing that the proposed method can be extended by adopting two alternate strategies of wavefront analysis to many classes of cells.

Breakthroughs in Photonics 2013: Holographic Imaging

Although holography is topic that goes back to the 1950s, the research in this field continues to be very active worldwide. A continuous growth is confirmed by the publication of more than 2000 papers each year in archival journal on different holographic issues. Here we describe shortly what appeared to us to be the most significant achievements reached in 2013 on holographic imaging.

Dynamic DIC by digital holography microscopy for enhancing phase-contrast visualization

Differential image contrast (DIC), through the numerical managing and manipulation of complex wavefronts obtained by digital holography (DH), is investigated. We name the approach Dynamical Differential Holographic Image Contrast (DDHIC). DDHIC dispenses from special optics and/or complex setup configurations with moveable components, as usually occurs in classical DIC, that is not well-suited for investigating objects experiencing dynamic evolution during the measurement. In fact, the technique presented here, is useful for floating samples since it allows, from a single recording, to set a posteriori the best conditions for DIC imaging in conjunction with the numerical focusing feature of DH. By DDHIC, the movies can be easily built-up to offering dynamic representation of phase-contrast along all directions, thus improving the visualization. Furthermore, the dynamic representation is useful for making the proper choice of other key parameters of DIC such as the amount of shear and the bias, with the aim to optimize the visualized phase-contrast imaging as favorite representation for bio-scientists. Investigation is performed on various biological samples.