Femtosecond laser fabricated monolithic chip for optical trapping and stretching of single cells.

We report on the fabrication by a femtosecond laser of an optofluidic device for optical trapping and stretching of single cells. Versatility and three-dimensional capabilities of this fabrication technology provide straightforward and extremely accurate alignment between the optical and fluidic components. Optical trapping and stretching of single red blood cells are demonstrated, thus proving the effectiveness of the proposed device as a monolithic optical stretcher. Our results pave the way for a new class of optofluidic devices for single cell analysis, in which, taking advantage of the flexibility of femtosecond laser micromachining, it is possible to further integrate sensing and sorting functions.

Design and optimization of a reflection-based fiber-optic tweezers.

We present the numerical modelling of a novel all-fiber optical tweezers, whose efficacy has been recently demonstrated. The device, realized by properly shaping the end-face of a fiber bundle, exploits total internal reflection to enhance the trapping efficiency. In order to allow the optimization of the performance, the trapping efficiency is evaluated as a function of different geometrical parameters of the structure. Given the peculiar spatial and angular distribution of the optical field, a new figure of merit is adopted to assess tweezers performance.

Kerr-induced spontaneous Bessel beam formation in the regime of strong two-photon absorption.

We study the effect of Two-Photon Absorption (TPA) nonlinear losses on Gaussian pulses, with power that exceeds the critical power for self-focusing, propagating in bulk kerr media. Experiments performed in fused silica and silicon highlight a spontaneous reshaping of the input pulse into a pulsed Bessel beam. A filament is formed in which sub-diffractive propagation is sustained by the Bessel-nature of the pulse.

Photodynamic action of merocyanine 540 on Staphylococcus epidermidis biofilms.

Photodynamic treatment (PDT) has been proposed as a new approach for inactivation of biofilms associated with medical devices that are resistant to chemical additives or biocides. In this study, we evaluated the antimicrobial activity of merocyanine 540 (MC 540), a photosensitizing dye that is used for purging malignant cells from autologous bone marrow grafts, against Staphylococcus epidermidis biofilms. Effect of the combined photodynamic action of MC 540 and 532 nm laser was investigated on the viability and structure of biofilms of two Staphylococcus epidermidis strains, RP62A and 1457. Significant inactivation of cells was observed when biofilms were exposed to MC 540 and laser simultaneously. The effect was found to be light dose-dependent but S. epidermidis 1457 biofilm proved to be slightly more susceptible than S. epidermidis RP62A biofilm. Furthermore, significant killing of both types of cells was attained even when a fixed light dose was delivered to the biofilms. Confocal laser scanning microscope (CLSM) analysis indicated damage to bacterial cell membranes in photodynamically treated biofilms, while disruption of PDT-treated biofilm was confirmed by scanning electron microscopy (SEM).

An integrated optofluidic device for single-cell sorting driven by mechanical properties.

We present a novel optofluidic device for real-time sorting on the basis of cell mechanical properties, measured by optical stretching. The whole mechanism, based on optical forces, does not hamper the viability of the tested cells, which can be used for further analysis. The device effectiveness is demonstrated by extracting a sample population enriched with highly metastatic cells from a heterogeneous cell mixture.

An optofluidic constriction chip for monitoring metastatic potential and drug response of cancer cells.

Cellular mechanical properties constitute good markers to characterize tumor cells, to study cell population heterogeneity and to highlight the effect of drug treatments. In this work, we describe the fabrication and validation of an integrated optofluidic chip capable of analyzing cellular deformability on the basis of the pressure gradient needed to push a cell through a narrow constriction. We demonstrate the ability of the chip to discriminate between tumorigenic and metastatic breast cancer cells (MCF7 and MDA-MB231) and between human melanoma cells with different metastatic potential (A375P and A375MC2). Moreover, we show that this chip allows highlighting the effect of drugs interfering with microtubule organization (paclitaxel, combretastatin A-4 and nocodazole) on cancer cells, which leads to changes in the pressure-gradient required to push cells through the constriction. Our single-cell microfluidic device for mechanical evaluation is compact and easy to use, allowing for an extensive use in different laboratory environments.

Effects of random and systematic perturbations in a one-dimensional photonic crystal wavelength converter.

We study the problem of the tolerance to fabrication errors in one-dimensional photonic crystal wavelength converters. In particular we consider the case of wavelength conversion obtained via quasiphase matching (QPM) based on a periodic amplitude modulation of the fundamental wave (Bloch-mode-QPM). Both numerical simulations of a waveguide-based structure and experimental results in an AlGaAs thin-film multilayer show that the proposed QPM mechanism is extremely tolerant to both systematic and random errors in the periodicity and duty cycle of the grating.

Integrated microfluidic device for single-cell trapping and spectroscopy.

Optofluidic microsystems are key components towards lab-on-a-chip devices for manipulation and analysis of biological specimens. In particular, the integration of optical tweezers (OT) in these devices allows stable sample trapping, while making available mechanical, chemical and spectroscopic analyses.

Optofluidic integrated cell sorter fabricated by femtosecond lasers.

The main trend in optofluidics is currently towards full integration of the devices, thus improving automation, compactness and portability. In this respect femtosecond laser microfabrication is a very powerful technology given its capability of producing both optical waveguides and microfluidic channels. The current challenge in biology is the possibility to perform bioassays at the single cell level to unravel the hidden complexity in nominally homogeneous populations. Here we report on a new device implementing a fully integrated fluorescence-activated cell sorter. This non-invasive device is specifically designed to operate with a limited amount of cells but with a very high selectivity in the sorting process. Characterization of the device with beads and validation with human cells are presented.

Complete retrieval of the field of ultrashort optical pulses using the angle-frequency spectrum.

We propose an experimental technique that allows for a complete characterization of the amplitude and phase of optical pulses in space and time. By the combination of a spatially resolved spectral measurement in the near and far fields and a frequency-resolved optical gating measurement, the electric field of the pulse is obtained through a fast, error-reduction algorithm.