Dynamics of mitochondrial membranes under photo-oxidative stress with high spatiotemporal resolution.

In our study, we harnessed an original Enhanced Speed Structured Illumination Microscopy (Fast-SIM) imaging setup to explore the dynamics of mitochondrial and inner membrane ultrastructure under specific photo-oxidation stress induced by Chlorin-e6 and light irradiation. Notably, our Fast-SIM system allowed us to observe and quantify a distinct remodeling and shortening of the mitochondrial structure after 60-80 s of irradiation. These changes were accompanied by fusion events of adjacent inner membrane cristae and global swelling of the organelle.

Enhanced neuroimaging with a calcium sensor in brains using closed-loop adaptive optics light-sheet fluorescence microscopy.

Significance: Adaptive optics (AO) has been implemented on several microscopy setups and has proven its ability to increase both signal and resolution. However, reported configurations are not suited for fast imaging of live samples or are based on an invasive or complex implementation method.

Aim: Provide a fast aberration correction method with an easy to implement AO module compatible with light-sheet fluorescence microscopy (LSFM) for enhanced imaging of live samples.

Adaptive optics light-sheet microscopy based on direct wavefront sensing without any guide star.

We propose an adaptive optics light-sheet fluorescence microscope (AO-LSFM) for closed-loop aberrations' correction at the emission path, providing intrinsic instrumental simplicity and high accuracy when compared to previously reported schemes. The approach is based on direct wavefront sensing, i.e.

In Vivo Imaging of Single Tumor Cells in Fast-Flowing Bloodstream Using Near-Infrared Quantum Dots and Time-Gated Imaging.

Whereas in vivo fluorescence imaging of cells immobilized within tissues provides a valuable tool to a broad range of biological studies, it still lacks the sensitivity required to visualize isolated cells circulating fast in the bloodstream due, in particular, to the autofluorescence from endogenous fluorophores. Time-gated imaging of near-infrared emitting ZnCuInSe/ZnS quantum dots (QDs) with fluorescence lifetimes in the range of 150-300 ns enables the efficient rejection of fast autofluorescence photons and the selection of QD fluorescence photons, thus significantly increasing sensitivity. We labeled model erythrocytes as well as lymphoma cells using these QDs coated with a stable zwitterionic polymer surface chemistry.

Sensorless adaptive optics implementation in widefield optical sectioning microscopy inside in vivo Drosophila brain.

We present an implementation of a sensorless adaptive optics loop in a widefield fluorescence microscope. This setup is designed to compensate for aberrations induced by the sample on both excitation and emission pathways. It allows fast optical sectioning inside a living Drosophila brain.

Oriented Bioconjugation of Unmodified Antibodies to Quantum Dots Capped with Copolymeric Ligands as Versatile Cellular Imaging Tools.

Distinctive optical properties of inorganic quantum dot (QD) nanoparticles promise highly valuable probes for fluorescence-based detection methods, particularly for in vivo diagnostics, cell phenotyping via multiple markers or single molecule tracking. However, despite high hopes, this promise has not been fully realized yet, mainly due to difficulties at producing stable, nontoxic QD bioconjugates of negligible nonspecific binding. Here, a universal platform for antibody binding to QDs is presented that builds upon the controlled functionalization of CdSe/CdS/ZnS nanoparticles capped with a multidentate dithiol/zwitterion copolymer ligand.

Time-gated cell imaging using long lifetime near-infrared-emitting quantum dots for autofluorescence rejection.

Fluorescence imaging is a promising technique for the detection of individual cell migration. Its sensitivity is, however, limited by a high tissue autofluorescence and a poor visible light penetration depth. In order to solve this problem, the fluorescence signal peak wavelength should lie in an absorption and diffusion free region and should be distinguishable, either spectrally or temporally, from the autofluorescence background.

Spectroscopy of single CdSe nanoplatelets.

We collect and resolve spectrally and temporally the photoluminescence of single CdSe nanoplatelets. The emission intensity of single nanoplatelets at room temperature shows ON and OFF periods with a usual blinking statistics, while at 20 K, their emission intensity can be extremely stable in time. At room temperature, the emission spectra of single nanoplatelets are similar to ensemble measurements with a full width at half-maximum of 40 meV.

Adaptive optics for fluorescence wide-field microscopy using spectrally independent guide star and markers.

We describe the implementation and use of an adaptive optics loop in the imaging path of a commercial wide field microscope. We show that it is possible to maintain the optical performances of the original microscope when imaging through aberrant biological samples. The sources used for illuminating the adaptive optics loop are spectrally independent, in excitation and emission, from the sample, so they do not appear in the final image, and their use does not contribute to the sample bleaching.

Bayesian estimation for optimized structured illumination microscopy.

Structured illumination microscopy is a recent imaging technique that aims at going beyond the classical optical resolution by reconstructing high-resolution (HR) images from low-resolution (LR) images acquired through modulation of the transfer function of the microscope. The classical implementation has a number of drawbacks, such as requiring a large number of images to be acquired and parameters to be manually set in an ad-hoc manner that have, until now, hampered its wide dissemination. Here, we present a new framework based on a Bayesian inverse problem formulation approach that enables the computation of one HR image from a reduced number of LR images and has no specific constraints on the modulation.

Single-shot optical sectioning using two-color probes in HiLo fluorescence microscopy.

We describe a wide-field fluorescence microscope setup which combines HiLo microscopy technique with the use of a two-color fluorescent probe. It allows one-shot fluorescence optical sectioning of thick biological moving sample which is illuminated simultaneously with a flat and a structured pattern at two different wavelengths. Both homogenous and structured fluorescence images are spectrally separated at detection and combined similarly with the HiLo microscopy technique.

Axial coding in full-field microscopy using three-dimensional structured illumination implemented with no moving parts.

We report a simple optical setup to produce both axial and lateral structured illumination through a single objective lens. With a minimum of six full-field images obtained without moving either the sample or the microscope objective, 100 nm diameter fluorescent beads can be localized axially with an accuracy of 50 nm in a 1.76-microm-thick layer.

Measurement of the optical parameters of the Virgo interferometer.

The Virgo interferometer, aimed at detecting gravitational waves, is now in a commissioning phase. Measurements of its optical properties are needed for the understanding of the instrument. We present the techniques developed for the measurement of the optical parameters of Virgo.

Confocal thermal-lens microscope.

The use of a confocal detection scheme in a dual-beam thermal-lens microscope is presented. The scheme allows the measurement of absorption factors down to 1.2 x 10(-7) in a 0.

Full-field birefringence imaging by thermal-light polarization-sensitive optical coherence tomography. II. Instrument and results.

We describe an instrument for measuring the magnitude of birefringence of tomographic images and the principal directions of axes that use thermal-light polarization-sensitive optical coherence tomography. The instrument permits full-field measurements with an axial resolution of 1.5 microm and a transverse resolution limited by diffraction.

Full-field birefringence imaging by thermal-light polarization-sensitive optical coherence tomography. I. Theory.

A method for measuring birefringence by use of thermal-light polarization-sensitive optical coherence tomography is presented. The use of thermal light brings to polarization-sensitive optical coherence tomography a resolution in the micrometer range in three dimensions. The instrument is based on a Linnik interference microscope and makes use of achromatic quarter-wave plates.

Absorption of low-loss optical materials measured at 1064 nm by a position-modulated collinear photothermal detection technique.

A collinear photothermal detection bench is described that makes use of a position-modulated heating source instead of the classic power-modulated source. This new modulation scheme increases by almost a factor 2 the sensitivity of a standard mirage bench. This bench is then used to measure the absorption coefficient of OH-free synthetic fused silica at 1064 nm in the parts per 10(6) range, which, combined with spectrophotometric measurements, confirms that the dominant absorption source is the OH content.