Tired and stressed: direct holographic quasi-static stretching of aging echinocytes and discocytes in plasma using optical tweezers [Invited].

Red blood cells (RBCs) undergo a progressive morphological transformation from smooth biconcave discocytes into rounder echinocytes with spicules on their surface during cold storage. The echinocytic morphology impacts RBCs' ability to flow through narrow sections of the circulation and therefore transfusion of RBC units with a high echinocytic content are thought to have a reduced efficiency. We use an optical tweezers-based technique where we directly trap and measure linear stiffness of RBCs under stress without the use of attached spherical probe particles or microfluidic flow to induce shear.

Roadmap on Deep Learning for Microscopy.

Through digital imaging, microscopy has evolved from primarily being a means for visual observation of life at the micro- and nano-scale, to a quantitative tool with ever-increasing resolution and throughput. Artificial intelligence, deep neural networks, and machine learning are all niche terms describing computational methods that have gained a pivotal role in microscopy-based research over the past decade. This Roadmap is written collectively by prominent researchers and encompasses selected aspects of how machine learning is applied to microscopy image data, with the aim of gaining scientific knowledge by improved image quality, automated detection, segmentation, classification and tracking of objects, and efficient merging of information from multiple imaging modalities.

Deep learning in light-matter interactions.

The deep-learning revolution is providing enticing new opportunities to manipulate and harness light at all scales. By building models of light-matter interactions from large experimental or simulated datasets, deep learning has already improved the design of nanophotonic devices and the acquisition and analysis of experimental data, even in situations where the underlying theory is not sufficiently established or too complex to be of practical use. Beyond these early success stories, deep learning also poses several challenges.

Enhanced Signal-to-Noise and Fast Calibration of Optical Tweezers Using Single Trapping Events.

The trap stiffness us the key property in using optical tweezers as a force transducer. Force reconstruction via maximum-likelihood-estimator analysis (FORMA) determines the optical trap stiffness based on estimation of the particle velocity from statistical trajectories. Using a modification of this technique, we determine the trap stiffness for a two micron particle within 2 ms to a precision of ∼10% using camera measurements at 10 kfps with the contribution of pixel noise to the signal being larger the level Brownian motion.

Strong Transient Flows Generated by Thermoplasmonic Bubble Nucleation.

The challenge of inducing and controlling localized fluid flows for generic force actuation and for achieving efficient mass transport in microfluidics is key to the development of next-generation miniaturized systems for chemistry and life sciences. Here we demonstrate a methodology for the robust generation and precise quantification of extremely strong flow transients driven by vapor bubble nucleation on spatially isolated plasmonic nanoantennas excited by light. The system is capable of producing peak flow speeds of the order mm/s at modulation rates up to ∼100 Hz in water, thus allowing for a variety of high-throughput applications.

Sound generation in zebrafish with Bio-Opto-Acoustics.

Hearing is a crucial sense in underwater environments for communication, hunting, attracting mates, and detecting predators. However, the tools currently used to study hearing are limited, as they cannot controllably stimulate specific parts of the auditory system. To date, the contributions of hearing organs have been identified through lesion experiments that inactivate an organ, making it difficult to gauge the specific stimuli to which each organ is sensitive, or the ways in which inputs from multiple organs are combined during perception.

Quantitative Acoustic Models for Superfluid Circuits.

We experimentally realize a highly tunable superfluid oscillator circuit in a quantum gas of ultracold atoms and develop and verify a simple lumped-element description of this circuit. At low oscillator currents, we demonstrate that the circuit is accurately described as a Helmholtz resonator, a fundamental element of acoustic circuits. At larger currents, the breakdown of the Helmholtz regime is heralded by a turbulent shedding of vortices and density waves.

Optical-trapping of particles in air using parabolic reflectors and a hollow laser beam.

We present an advanced optical-trapping method that is capable of trapping arbitrary shapes of transparent and absorbing particles in air. Two parabolic reflectors were used to reflect the inner and outer parts of a single hollow laser beam, respectively, to form two counter-propagating conical beams and bring them into a focal point for trapping. This novel design demonstrated high trapping efficiency and strong trapping robustness with a simple optical configuration.

Giant vortex clusters in a two-dimensional quantum fluid.

Adding energy to a system through transient stirring usually leads to more disorder. In contrast, point-like vortices in a bounded two-dimensional fluid are predicted to reorder above a certain energy, forming persistent vortex clusters. In this study, we experimentally realize these vortex clusters in a planar superfluid: a Rb Bose-Einstein condensate confined to an elliptical geometry.

High-speed transverse and axial optical force measurements using amplitude filter masks.

Direct optical force measurement is a versatile method used in optical tweezers experiments, providing accurate measurements of forces for a wide range of particles and trapping beams. It is based on the detection of the change of the momentum of light scattered by a trapped object. A digital micromirror device can be used to selectively reflect light in different directions using an appropriately defined mask.

Measuring local properties inside a cell-mimicking structure using rotating optical tweezers.

Exploring the rheological properties of intracellular materials is essential for understanding cellular and subcellular processes. Optical traps have been widely used for physical manipulation of micro and nano objects within fluids enabling studies of biological systems. However, experiments remain challenging as it is unclear how the probe particle's mobility is influenced by the nearby membranes and organelles.

Cellular-Resolution Imaging of Vestibular Processing across the Larval Zebrafish Brain.

The vestibular system, which reports on motion and gravity, is essential to postural control, balance, and egocentric representations of movement and space. The motion needed to stimulate the vestibular system complicates studying its circuitry, so we previously developed a method for fictive vestibular stimulation in zebrafish, using optical trapping to apply physical forces to the otoliths. Here, we combine this approach with whole-brain calcium imaging at cellular resolution, delivering a comprehensive map of the brain regions and cellular responses involved in basic vestibular processing.

Calibration of force detection for arbitrarily shaped particles in optical tweezers.

Force measurement with an optical trap requires calibration of it. With a suitable detector, such as a position-sensitive detector (PSD), it is possible to calibrate the detector so that the force can be measured for arbitrary particles and arbitrary beams without further calibration; such a calibration can be called an "absolute calibration". Here, we present a simple method for the absolute calibration of a PSD.

Modelling of Cavity Optomechanical Magnetometers.

Cavity optomechanical magnetic field sensors, constructed by coupling a magnetostrictive material to a micro-toroidal optical cavity, act as ultra-sensitive room temperature magnetometers with tens of micrometre size and broad bandwidth, combined with a simple operating scheme. Here, we develop a general recipe for predicting the field sensitivity of these devices. Several geometries are analysed, with a highest predicted sensitivity of 180 p T / Hz at 28 μ m resolution limited by thermal noise in good agreement with previous experimental observations.

Hypothalamic Projections to the Optic Tectum in Larval Zebrafish.

The optic tectum of larval zebrafish is an important model for understanding visual processing in vertebrates. The tectum has been traditionally viewed as dominantly visual, with a majority of studies focusing on the processes by which tectal circuits receive and process retinally-derived visual information. Recently, a handful of studies have shown a much more complex role for the optic tectum in larval zebrafish, and anatomical and functional data from these studies suggest that this role extends beyond the visual system, and beyond the processing of exclusively retinal inputs.

Impact of complex surfaces on biomicrorheological measurements using optical tweezers.

The characterisation of physical properties in biologically relevant processes and the development of novel microfluidic devices for this purpose are experiencing a great resurgence at present. In many of measurements of this type where a probe in a fluid is used, the strong influence of the boundaries of the volume used is a serious problem. In these geometries the proximity of a probe to a wall can severely influence the measurement.

Rotational control of computer generated holograms.

We develop a basis for three-dimensional rotation of arbitrary light fields created by computer generated holograms. By adding an extra phase function into the kinoform, any light field or holographic image can be tilted in the focal plane with minimized distortion. We present two different approaches to rotate an arbitrary hologram: the Scheimpflug method and a novel coordinate transformation method.

Optical trapping of otoliths drives vestibular behaviours in larval zebrafish.

The vestibular system, which detects gravity and motion, is crucial to survival, but the neural circuits processing vestibular information remain incompletely characterised. In part, this is because the movement needed to stimulate the vestibular system hampers traditional neuroscientific methods. Optical trapping uses focussed light to apply forces to targeted objects, typically ranging from nanometres to a few microns across.

Active rotational and translational microrheology beyond the linear spring regime.

Active particle tracking microrheometers have the potential to perform accurate broadband measurements of viscoelasticity within microscopic systems. Generally, their largest possible precision is limited by Brownian motion and low frequency changes to the system. The signal to noise ratio is usually improved by increasing the size of the driven motion compared to the Brownian as well as averaging over repeated measurements.

Nondestructive Profilometry of Optical Nanofibers.

Single-mode optical nanofibers are a central component of a broad range of applications and emerging technologies. Their fabrication has been extensively studied over the past decade, but imaging of the final submicrometer products has been restricted to destructive or low-precision techniques. Here, we demonstrate an optical scattering-based scanning method that uses a probe nanofiber to locally scatter the evanescent field of a sample nanofibre.