Heating on the microscale using focused lasers gave rise to recent applications, e.g., in biomedicine, biology and microfluidics, especially using gold nanoparticles as efficient nanoabsorbers of light.
View Article and Find Full Text PDFPhase and fluorescence are complementary contrasts that are commonly used in biology. However, the coupling of these two modalities is traditionally limited to high magnification and complex imaging systems. For statistical studies of biological populations, a large field-of-view is required.
View Article and Find Full Text PDFSingle-molecule localization microscopy (SMLM) enables the production of high-resolution images by imaging spatially isolated fluorescent particles. Although challenging, the result of SMLM analysis lists the position of individual molecules, leading to a valuable quantification of the stoichiometry and spatial organization of molecular actors. Both the signal/noise ratio and the density (D), i.
View Article and Find Full Text PDFLaser heating of individual cells in culture recently led to seminal studies in cell poration, fusion, migration, or nanosurgery, although measuring the local temperature increase in such experiments remains a challenge. Here, the laser-induced dynamical control of the heat-shock response is demonstrated at the single cell level, enabled by the use of light-absorbing gold nanoparticles as nanosources of heat and a temperature mapping technique based on quadriwave lateral shearing interferometry (QLSI) measurements. As it is label-free, this approach does not suffer from artifacts inherent to previously reported fluorescence-based temperature-mapping techniques and enables the use of any standard fluorescent labels to monitor in parallel the cell's response.
View Article and Find Full Text PDFA technique that provides quantitative and spatially resolved retardance measurement is studied for application to laser-induced modification in transparent materials. The method is based on the measurement of optical path differences between two wavefronts carrying different polarizations, measured by a wavefront sensor placed in the image plane of a microscope. We have applied the technique to the investigation of stress distribution induced by CO laser processing of fused silica samples.
View Article and Find Full Text PDFWe report on a simple and efficient technique based on a wavefront sensor to obtain time-resolved amplitude and phase images of laser-material interactions. The main interest of the technique is to obtain quantitative self-calibrated phase measurements in one shot at the femtosecond time-scale, with high spatial resolution. The technique is used for direct observation and quantitative measurement of the Kerr effect in a fused silica substrate and free electron generation by photo-ionization processes in an optical coating.
View Article and Find Full Text PDFSolvothermal synthesis, denoting chemical reactions occurring in metastable liquids above their boiling point, normally requires the use of a sealed autoclave under pressure to prevent the solvent from boiling. This work introduces an experimental approach that enables solvothermal synthesis at ambient pressure in an open reaction medium. The approach is based on the use of gold nanoparticles deposited on a glass substrate and acting as photothermal sources.
View Article and Find Full Text PDFRigid endoscopes like graded-index (GRIN) lenses are known tools in biological imaging, but it is conceptually difficult to miniaturize them. In this letter, we demonstrate an ultra-thin rigid endoscope with a diameter of only 125 μm. In addition, we identify a domain where two-photon endoscopic imaging with fs-pulse excitation is possible.
View Article and Find Full Text PDFEssential cellular functions as diverse as genome maintenance and tissue morphogenesis rely on the dynamic organization of filamentous assemblies. For example, the precise structural organization of DNA filaments has profound consequences on all DNA-mediated processes including gene expression, whereas control over the precise spatial arrangement of cytoskeletal protein filaments is key for mechanical force generation driving animal tissue morphogenesis. Polarized fluorescence is currently used to extract structural organization of fluorescently labeled biological filaments by determining the orientation of fluorescent labels, however with a strong drawback: polarized fluorescence imaging is indeed spatially limited by optical diffraction, and is thus unable to discriminate between the intrinsic orientational mobility of the fluorophore labels and the real structural disorder of the labeled biomolecules.
View Article and Find Full Text PDFSingle-cell dry mass measurement is used in biology to follow cell cycle, to address effects of drugs, or to investigate cell metabolism. Quantitative phase imaging technique with quadriwave lateral shearing interferometry (QWLSI) allows measuring cell dry mass. The technique is very simple to set up, as it is integrated in a camera-like instrument.
View Article and Find Full Text PDFWe investigate phase imaging as a measurement method for laser damage detection and analysis of laser-induced modification of optical materials. Experiments have been conducted with a wavefront sensor based on lateral shearing interferometry associated with a high-magnification optical microscope. The system has been used for the in-line observation of optical thin films and bulk samples, laser irradiated in two different conditions: 500 fs pulses at 343 and 1030 nm, and millisecond to second irradiation with a CO2 laser at 10.
View Article and Find Full Text PDFMeeting the nanometre resolution promised by super-resolution microscopy techniques (pointillist: PALM, STORM, scanning: STED) requires stabilizing the sample drifts in real time during the whole acquisition process. Metal nanoparticles are excellent probes to track the lateral drifts as they provide crisp and photostable information. However, achieving nanometre axial super-localization is still a major challenge, as diffraction imposes large depths-of-fields.
View Article and Find Full Text PDFWe describe a new technique based on the use of a high-resolution quadri-wave lateral shearing interferometer to perform quantitative linear retardance and birefringence measurements on biological samples. The system combines quantitative phase images with varying polarization excitation to create retardance images. This technique is compatible with living samples and gives information about the local retardance and structure of their anisotropic components.
View Article and Find Full Text PDFWe introduce a deterministic procedure, named TSUNA for Temperature Shaping Using Nanoparticle Assemblies, aimed at generating arbitrary temperature distributions on the microscale. The strategy consists in (i) using an inversion algorithm to determine the exact heat source density necessary to create a desired temperature distribution and (ii) reproducing experimentally this calculated heat source density using smart assemblies of lithographic metal nanoparticles under illumination at their plasmonic resonance wavelength. The feasibility of this approach was demonstrated experimentally by thermal microscopy based on wavefront sensing.
View Article and Find Full Text PDFTo investigate the early stages of cell-cell interactions occurring between living biological samples, imaging methods with appropriate spatiotemporal resolution are required. Among the techniques currently available, those based on optical trapping are promising. Methods to image trapped objects, however, in general suffer from a lack of three-dimensional resolution, due to technical constraints.
View Article and Find Full Text PDFWe describe the use of spatially incoherent illumination to make quantitative phase imaging of a semi-transparent sample, even out of the paraxial approximation. The image volume electromagnetic field is collected by scanning the image planes with a quadriwave lateral shearing interferometer, while the sample is spatially incoherently illuminated. In comparison to coherent quantitative phase measurements, incoherent illumination enriches the 3D collected spatial frequencies leading to 3D resolution increase (up to a factor 2).
View Article and Find Full Text PDFWe report a first demonstration of two-photon endoscopic imaging with a lensless endoscope. The endoscope probe is a double-clad bundle of single-mode fibers; point excitation and scanning is achieved by coherent combining of femtosecond light pulses propagating in the single-mode fibers; and back-scattered two-photon signal is collected through the multi-mode inner cladding. We demonstrate the two-photon endoscope on a test sample of rhodamine 6G crystals.
View Article and Find Full Text PDFThe temperature distribution throughout arrays of illuminated metal nanoparticles is investigated numerically and experimentally. The two cases of continuous and femtosecond-pulsed illumination are addressed. In the case of continuous illumination, two distinct regimes are evidenced: a temperature confinement regime, where the temperature increase remains confined at the vicinity of each nanosource of heat, and a temperature delocalization regime, where the temperature is uniform throughout the whole nanoparticle assembly despite the heat sources' nanometric size.
View Article and Find Full Text PDFWe present a method that allows one to measure the real and imaginary parts of the third-order susceptibility in a wide-field coherent anti-Stokes Raman scattering setup using a quadriwave lateral shearing interferometer. This permits the retrieval of the undistorted Raman spectrum and the removal of a nonresonant signal from the surrounding solvent, which otherwise may overwhelm weak resonances.
View Article and Find Full Text PDFWe report a step toward scanning endomicroscopy without distal optics. The focusing of the beam at the distal end of a fiber bundle is achieved by imposing a parabolic phase profile across the exit face with the aid of a spatial light modulator. We achieve video-rate images by galvanometric scanning of the phase tilt at the proximal end.
View Article and Find Full Text PDFWe propose and implement a wide-field microscopy method to retrieve the real and imaginary part of a field emitted by coherent and resonant molecular scatterers. The technique is based on wave-front sensing and does not require the use of any reference beam. We exemplify its ability in wide-field coherent anti-Stokes Raman scattering imaging and retrieve the complex anti-Stokes field while spectrally scanning a molecular vibrational resonance.
View Article and Find Full Text PDFA wavefront sensor is used as a direct observation tool to image the Gouy phase shift in photonic nanojets created by micrometer-sized dielectric spheres. The amplitude and phase distributions of light are found in good agreement with a rigorous electromagnetic computation. Interestingly the observed phase shift when travelling through the photonic jet is a combination of the awaited π Gouy shift and a phase shift induced by the bead refraction.
View Article and Find Full Text PDFWe present a quadriwave lateral shearing interferometer used as a wavefront sensor and mounted on a commercial non-modified transmission white-light microscope as a quantitative phase imaging technique. The setup is designed to simultaneously make measurements with both quantitative transmission phase and fluorescence modes: phase enables enhanced contrasted visualization of the cell structure including intracellular organelles, while fluorescence allows a complete and precise identification of each component. After the characterization of the phase measurement reliability and sensitivity on calibrated samples, we use these two imaging modes to measure the characteristic optical path difference between subcellular elements (mitochondria, actin fibers, and vesicles) and cell medium, and demonstrate that phase-only information should be sufficient to identify some organelles without any labeling, like lysosomes.
View Article and Find Full Text PDFWavefront sensors are usually based on measuring the wavefront derivatives. The most commonly used approach to quantitatively reconstruct the wavefront uses discrete Fourier transform, which leads to artifacts when phase objects are located at the image borders. We propose here a simple approach to avoid these artifacts based on the duplication and antisymmetrization of the derivatives data, in the derivative direction, before integration.
View Article and Find Full Text PDFA generalized product-of-convolution model for simulation of quantitative phase microscopy of thick heterogeneous specimen under tilted plane-wave illumination is presented. Actual simulations are checked against a much more time-consuming commercial finite-difference time-domain method. Then modeled data are compared with experimental measurements that were made with a quadriwave lateral shearing interferometer.
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