Enhanced Quantitative Wavefront Imaging for Nano-Object Characterization.

Quantitative phase imaging enables precise and label-free characterizations of individual nano-objects within a large volume, without a priori knowledge of the sample or imaging system. While emerging common path implementations are simple enough to promise a broad dissemination, their phase sensitivity still falls short of precisely estimating the mass or polarizability of vesicles, viruses, or nanoparticles in single-shot acquisitions. In this paper, we revisit the Zernike filtering concept, originally crafted for intensity-only detectors, with the aim of adapting it to wavefront imaging. We demonstrate, through numerical simulation and experiments based on high-resolution wavefront sensing, that a simple Fourier-plane add-on can significantly enhance phase sensitivity for subdiffraction objects─achieving over an order of magnitude increase (×12)─while allowing the quantitative retrieval of both intensity and phase. This advancement allows for more precise nano-object detection and metrology.

Life at high temperature observed in vitro upon laser heating of gold nanoparticles.

Thermophiles are microorganisms that thrive at high temperature. Studying them can provide valuable information on how life has adapted to extreme conditions. However, high temperature conditions are difficult to achieve on conventional optical microscopes. Some home-made solutions have been proposed, all based on local resistive electric heating, but no simple commercial solution exists. In this article, we introduce the concept of microscale laser heating over the field of view of a microscope to achieve high temperature for the study of thermophiles, while maintaining the user environment in soft conditions. Microscale heating with moderate laser intensities is achieved using a substrate covered with gold nanoparticles, as biocompatible, efficient light absorbers. The influences of possible microscale fluid convection, cell confinement and centrifugal thermophoretic motion are discussed. The method is demonstrated with two species: (i) Geobacillus stearothermophilus, a motile thermophilic bacterium thriving around 65 °C, which we observed to germinate, grow and swim upon microscale heating and (ii) Sulfolobus shibatae, a hyperthermophilic archaeon living at the optimal temperature of 80 °C. This work opens the path toward simple and safe observation of thermophilic microorganisms using current and accessible microscopy tools.

Fast photothermal spatial light modulation for quantitative phase imaging at the nanoscale.

Spatial light modulators have become an essential tool for advanced microscopy, enabling breakthroughs in 3D, phase, and super-resolution imaging. However, continuous spatial-light modulation that is capable of capturing sub-millisecond microscopic motion without diffraction artifacts and polarization dependence is challenging. Here we present a photothermal spatial light modulator (PT-SLM) enabling fast phase imaging for nanoscopic 3D reconstruction. The PT-SLM can generate a step-like wavefront change, free of diffraction artifacts, with a high transmittance and a modulation efficiency independent of light polarization. We achieve a phase-shift > π and a response time as short as 70 µs with a theoretical limit in the sub microsecond range. We used the PT-SLM to perform quantitative phase imaging of sub-diffractional species to decipher the 3D nanoscopic displacement of microtubules and study the trajectory of a diffusive microtubule-associated protein, providing insights into the mechanism of protein navigation through a complex microtubule network.

Microscale Temperature Shaping Using Spatial Light Modulation on Gold Nanoparticles.

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. However, such an approach naturally leads to nonuniform, Gaussian-like temperature distributions due to the diffusive nature of heat. Here, we report on an experimental means to generate arbitrary distributions of temperature profiles on the micrometric scale (e.g. uniform, linear, parabolic, etc) consisting in illuminating a uniform gold nanoparticle distribution on a planar substrate using spatially contrasted laser beams, shaped using a spatial light modulator (SLM). We explain how to compute the light pattern and the SLM interferogram to achieve the desired temperature distribution, and demonstrate the approach by carrying out temperature measurements using quantitative wavefront sensing.

Photothermal Control of Heat-Shock Protein Expression at the Single Cell Level.

Laser 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.

Light-Assisted Solvothermal Chemistry Using Plasmonic Nanoparticles.

Solvothermal 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. To illustrate the approach, the selected hydrothermal reaction involves the formation of indium hydroxide microcrystals favored at 200 °C in liquid water. In addition to demonstrating the principle, the benefits and the specific characteristics of such an approach are investigated, in particular, the much faster reaction rate, the achievable spatial and time scales, the effect of microscale temperature gradients, the effect of the size of the heated area, and the effect of thermal-induced microscale fluid convection. This technique is general and could be used to spatially control the deposition of virtually any material for which a solvothermal synthesis exists.