Label-free optical imaging employs natural and nondestructive approaches for the visualisation of biomedical samples for both biological assays and clinical diagnosis. Currently, this field revolves around multiple broad technology-oriented communities, each with a specific focus on a particular modality despite the existence of shared challenges and applications. As a result, biologists or clinical researchers who require label-free imaging are often not aware of the most appropriate modality to use. This manuscript presents a comprehensive review of and comparison among different label-free imaging modalities and discusses common challenges and applications. We expect this review to facilitate collaborative interactions between imaging communities, push the field forward and foster technological advancements, biophysical discoveries, as well as clinical detection, diagnosis, and monitoring of disease.

Bioimaging harnessing optical contrasts and chemical specificity is of vital importance in probing complex biology. Vibrational spectroscopy based on mid-infrared (mid-IR) excitation can reveal rich chemical information about molecular distributions. However, its full potential for bioimaging is hindered by the achievable sensitivity. Here, we report bond selective fluorescence-detected infrared-excited (BonFIRE) spectral microscopy. BonFIRE employs two-photon excitation in the mid-IR and near-IR to upconvert vibrational excitations to electronic states for fluorescence detection, thus encoding vibrational information into fluorescence. The system utilizes tuneable narrowband picosecond pulses to ensure high sensitivity, biocompatibility, and robustness for bond-selective biological interrogations over a wide spectrum of reporter molecules. We demonstrate BonFIRE spectral imaging in both fingerprint and cell-silent spectroscopic windows with single-molecule sensitivity for common fluorescent dyes. We then demonstrate BonFIRE imaging on various intracellular targets in fixed and live cells, neurons, and tissues, with promises for further vibrational multiplexing. For dynamic bioanalysis in living systems, we implement a high-frequency modulation scheme and demonstrate time-lapse BonFIRE microscopy of live HeLa cells. We expect BonFIRE to expand the bioimaging toolbox by providing a new level of bond-specific vibrational information and facilitate functional imaging and sensing for biological investigations.

Wide field of view microscopy that can resolve 3D information at high speed and spatial resolution is highly desirable for studying the behaviour of freely moving model organisms. However, it is challenging to design an optical instrument that optimises all these properties simultaneously. Existing techniques typically require the acquisition of sequential image snapshots to observe large areas or measure 3D information, thus compromising on speed and throughput. Here, we present 3D-RAPID, a computational microscope based on a synchronized array of 54 cameras that can capture high-speed 3D topographic videos over an area of 135 cm2, achieving up to 230 frames per second at spatiotemporal throughputs exceeding 5 gigapixels per second. 3D-RAPID employs a 3D reconstruction algorithm that, for each synchronized snapshot, fuses all 54 images into a composite that includes a co-registered 3D height map. The self-supervised 3D reconstruction algorithm trains a neural network to map raw photometric images to 3D topography using stereo overlap redundancy and ray-propagation physics as the only supervision mechanism. The resulting reconstruction process is thus robust to generalization errors and scales to arbitrarily long videos from arbitrarily sized camera arrays. We demonstrate the broad applicability of 3D-RAPID with collections of several freely behaving organisms, including ants, fruit flies, and zebrafish larvae.

Wavefront shaping (WFS) is emerging as a promising tool for controlling and focusing light in complex scattering media. The shaping system's speed, the energy gain of the corrected wavefronts, and the control degrees of freedom (DOF) are the most important metrics for WFS, especially for highly scattering and dynamic samples. Despite recent advances, current methods suffer from trade-offs that limit satisfactory performance to only one or two of these metrics. Here, we report a WFS technique that simultaneously achieves high speed, high energy gain, and high control DOF. By combining photorefractive crystal-based analog optical phase conjugation (AOPC) and stimulated emission light amplification, our technique achieves an energy gain approaching unity, more than three orders of magnitude larger than conventional AOPC. The response time of ~10 s with about 10 control modes corresponds to an average mode time of about 0.01 ns/mode, which is more than 50 times lower than some of the fastest WFS systems to date. We anticipate that this technique will be instrumental in overcoming the optical diffusion limit in photonics and translate WFS techniques to real-world applications.

Widefield microscopy of optically thick specimens typically features reduced contrast due to "spatial crosstalk", in which the signal at each point in the field of view is the result of a superposition from neighbouring points that are simultaneously illuminated. In 1955, Marvin Minsky proposed confocal microscopy as a solution to this problem. Today, laser scanning confocal fluorescence microscopy is broadly used due to its high depth resolution and sensitivity, but comes at the price of photobleaching, chemical, and photo-toxicity. Here, we present artificial confocal microscopy (ACM) to achieve confocal-level depth sectioning, sensitivity, and chemical specificity, on unlabeled specimens, nondestructively. We equipped a commercial laser scanning confocal instrument with a quantitative phase imaging module, which provides optical path-length maps of the specimen in the same field of view as the fluorescence channel. Using pairs of phase and fluorescence images, we trained a convolution neural network to translate the former into the latter. The training to infer a new tag is very practical as the input and ground truth data are intrinsically registered, and the data acquisition is automated. The ACM images present significantly stronger depth sectioning than the input (phase) images, enabling us to recover confocal-like tomographic volumes of microspheres, hippocampal neurons in culture, and 3D liver cancer spheroids. By training on nucleus-specific tags, ACM allows for segmenting individual nuclei within dense spheroids for both cell counting and volume measurements. In summary, ACM can provide quantitative, dynamic data, nondestructively from thick samples, while chemical specificity is recovered computationally.

Imaging both the positions and orientations of single fluorophores, termed single-molecule orientation-localisation microscopy, is a powerful tool to study biochemical processes. However, the limited photon budget associated with single-molecule fluorescence makes high-dimensional imaging with isotropic, nanoscale spatial resolution a formidable challenge. Here, we realise a radially and azimuthally polarized multi-view reflector (raMVR) microscope for the imaging of the 3D positions and 3D orientations of single molecules, with precision of 10.9 nm and 2.0° over a 1.5 μm depth range. The raMVR microscope achieves 6D super-resolution imaging of Nile red (NR) molecules transiently bound to lipid-coated spheres, accurately resolving their spherical morphology despite refractive-index mismatch. By observing the rotational dynamics of NR, raMVR images also resolve the infiltration of lipid membranes by amyloid-beta oligomers without covalent labelling. Finally, we demonstrate 6D imaging of cell membranes, where the orientations of specific fluorophores reveal heterogeneity in membrane fluidity. With its nearly isotropic 3D spatial resolution and orientation measurement precision, we expect the raMVR microscope to enable 6D imaging of molecular dynamics within biological and chemical systems with exceptional detail.

Optical-resolution photoacoustic microscopy (OR-PAM) can visualize wavelength-dependent optical absorption at the cellular level. However, OR-PAM suffers from a limited depth of field (DOF) due to the tight focus of the optical excitation beam, making it challenging to acquire high-resolution images of samples with uneven surfaces or high-quality volumetric images without z-scanning. To overcome this limitation, we propose needle-shaped beam photoacoustic microscopy (NB-PAM), which can extend the DOF to up to ~28-fold Rayleigh lengths via customized diffractive optical elements (DOEs). The DOE generate a needle beam with a well-maintained beam diameter, a uniform axial intensity distribution, and negligible sidelobes. The advantage of using NB-PAM is demonstrated by both histology-like imaging of fresh slide-free organs using a 266 nm laser and in vivo mouse brain vasculature imaging using a 532 nm laser. The approach provides new perspectives for slide-free intraoperative pathological imaging and organ-level imaging.

Quantitative Phase Imaging (QPI) has gained popularity in bioimaging because it can avoid the need for cell staining, which in some cases is difficult or impossible. However, as a result, QPI does not provide labelling of various specific intracellular structures. Here we show a novel computational segmentation method based on statistical inference that makes it possible for QPI techniques to identify the cell nucleus. We demonstrate the approach with refractive index tomograms of stain-free cells reconstructed through the tomographic phase microscopy in flow cytometry mode. In particular, by means of numerical simulations and two cancer cell lines, we demonstrate that the nucleus can be accurately distinguished within the stain-free tomograms. We show that our experimental results are consistent with confocal fluorescence microscopy (FM) data and microfluidic cytofluorimeter outputs. This is a significant step towards extracting specific three-dimensional intracellular structures directly from the phase-contrast data in a typical flow cytometry configuration.

Microscopic imaging in three dimensions enables numerous biological and clinical applications. However, high-resolution optical imaging preserved in a relatively large depth range is hampered by the rapid spread of tightly confined light due to diffraction. Here, we show that a particular disposition of light illumination and collection paths liberates optical imaging from the restrictions imposed by diffraction. This arrangement, realized by metasurfaces, decouples lateral resolution from depth-of-focus by establishing a one-to-one correspondence (bijection) along a focal line between the incident and collected light. Implementing this approach in optical coherence tomography, we demonstrate tissue imaging at 1.3 μm wavelength with ~ 3.2 μm lateral resolution, maintained nearly intact over 1.25 mm depth-of-focus, with no additional acquisition or computation burden. This method, termed bijective illumination collection imaging, is general and might be adapted across various existing imaging modalities.

X-ray and gamma-ray photons are widely used for imaging but require a mathematical reconstruction step, known as tomography, to produce cross-sectional images from the measured data. Theoretically, the back-to-back annihilation photons produced by positron-electron annihilation can be directly localized in three-dimensional space using time-of-flight information without tomographic reconstruction. However, this has not yet been demonstrated due to the insufficient timing performance of available radiation detectors. Here, we develop techniques based on detecting prompt Cerenkov photons, which when combined with a convolutional neural network for timing estimation resulted in an average timing precision of 32 picoseconds, corresponding to a spatial precision of 4.8 mm. We show this is sufficient to produce cross-sectional images of a positron-emitting radionuclide directly from the detected coincident annihilation photons, without using any tomographic reconstruction algorithm. The reconstruction-free imaging demonstrated here directly localizes positron emission, and frees the design of an imaging system from the geometric and sampling constraints that normally present for tomographic reconstruction.

Optical information processing and computing can potentially offer enhanced performance, scalability and energy efficiency. However, achieving nonlinearity-a critical component of computation-remains challenging in the optical domain. Here we introduce a design that leverages a multiple-scattering cavity to passively induce optical nonlinear random mapping with a continuous-wave laser at a low power. Each scattering event effectively mixes information from different areas of a spatial light modulator, resulting in a highly nonlinear mapping between the input data and output pattern. We demonstrate that our design retains vital information even when the readout dimensionality is reduced, thereby enabling optical data compression. This capability allows our optical platforms to offer efficient optical information processing solutions across applications. We demonstrate our design's efficacy across tasks, including classification, image reconstruction, keypoint detection and object detection, all of which are achieved through optical data compression combined with a digital decoder. In particular, high performance at extreme compression ratios is observed in real-time pedestrian detection. Our findings open pathways for novel algorithms and unconventional architectural designs for optical computing.

The topological response of matter to electromagnetic fields is a highly demanded property in materials design and metrology due to its robustness against noise and decoherence, stimulating recent advances in ultrafast photonics. Embedding topological properties into the enantiosensitive optical response of chiral molecules could therefore enhance the efficiency and robustness of chiral optical discrimination. Here we achieve such a topological embedding by introducing the concept of chiral topological light-a light beam which displays chirality locally, with an azimuthal distribution of its handedness described globally by a topological charge. The topological charge is mapped onto the azimuthal intensity modulation of the non-linear optical response, where enantiosensitivity is encoded into its spatial rotation. The spatial rotation is robust against intensity fluctuations and imperfect local polarization states of the driving field. Our theoretical results show that chiral topological light enables detection of percentage-level enantiomeric excesses in randomly oriented mixtures of chiral molecules, opening a way to new, extremely sensitive and robust chiro-optical spectroscopies with attosecond time resolution.

Extreme miniaturization of infrared spectrometers is critical for their integration into next-generation consumer electronics, wearables and ultrasmall satellites. In the infrared, there is a necessary compromise between high spectral bandwidth and high spectral resolution when miniaturizing dispersive elements, narrow band-pass filters and reconstructive spectrometers. Fourier-transform spectrometers are known for their large bandwidth and high spectral resolution in the infrared; however, they have not been fully miniaturized. Waveguide-based Fourier-transform spectrometers offer a low device footprint, but rely on an external imaging sensor such as bulky and expensive InGaAs cameras. Here we demonstrate a proof-of-concept miniaturized Fourier-transform waveguide spectrometer that incorporates a subwavelength and complementary-metal-oxide-semiconductor-compatible colloidal quantum dot photodetector as a light sensor. The resulting spectrometer exhibits a large spectral bandwidth and moderate spectral resolution of 50 cm at a total active spectrometer volume below 100 μm × 100 μm × 100 μm. This ultracompact spectrometer design allows the integration of optical/analytical measurement instruments into consumer electronics and space devices.

The development of luminescent organic radicals has resulted in materials with excellent optical properties for near-infrared emission. Applications of light generation in this range span from bioimaging to surveillance. Although the unpaired electron arrangements of radicals enable efficient radiative transitions within the doublet-spin manifold in organic light-emitting diodes, their performance is limited by non-radiative pathways introduced in electroluminescence. Here we present a host-guest design for organic light-emitting diodes that exploits energy transfer with up to 9.6% external quantum efficiency for 800 nm emission. The tris(2,4,6-trichlorophenyl)methyl-triphenyl-amine radical guest is energy-matched to the triplet state in a charge-transporting anthracene-derivative host. We show from optical spectroscopy and quantum-chemical modelling that reversible host-guest triplet-doublet energy transfer allows efficient harvesting of host triplet excitons.

Laser frequency combs are enabling some of the most exciting scientific endeavours in the twenty-first century, ranging from the development of optical clocks to the calibration of the astronomical spectrographs used for discovering Earth-like exoplanets. Dissipative Kerr solitons generated in microresonators currently offer the prospect of attaining frequency combs in miniaturized systems by capitalizing on advances in photonic integration. Most of the applications based on soliton microcombs rely on tuning a continuous-wave laser into a longitudinal mode of a microresonator engineered to display anomalous dispersion. In this configuration, however, nonlinear physics precludes one from attaining dissipative Kerr solitons with high power conversion efficiency, with typical comb powers amounting to ~1% of the available laser power. Here we demonstrate that this fundamental limitation can be overcome by inducing a controllable frequency shift to a selected cavity resonance. Experimentally, we realize this shift using two linearly coupled anomalous-dispersion microresonators, resulting in a coherent dissipative Kerr soliton with a conversion efficiency exceeding 50% and excellent line spacing stability. We describe the soliton dynamics in this configuration and find vastly modified characteristics. By optimizing the microcomb power available on-chip, these results facilitate the practical implementation of a scalable integrated photonic architecture for energy-efficient applications.

Lightning discharges between charged clouds and the Earth's surface are responsible for considerable damages and casualties. It is therefore important to develop better protection methods in addition to the traditional Franklin rod. Here we present the first demonstration that laser-induced filaments-formed in the sky by short and intense laser pulses-can guide lightning discharges over considerable distances. We believe that this experimental breakthrough will lead to progress in lightning protection and lightning physics. An experimental campaign was conducted on the Säntis mountain in north-eastern Switzerland during the summer of 2021 with a high-repetition-rate terawatt laser. The guiding of an upward negative lightning leader over a distance of 50 m was recorded by two separate high-speed cameras. The guiding of negative lightning leaders by laser filaments was corroborated in three other instances by very-high-frequency interferometric measurements, and the number of X-ray bursts detected during guided lightning events greatly increased. Although this research field has been very active for more than 20 years, this is the first field-result that experimentally demonstrates lightning guided by lasers. This work paves the way for new atmospheric applications of ultrashort lasers and represents an important step forward in the development of a laser based lightning protection for airports, launchpads or large infrastructures.

Entanglement between particles is a basic concept of quantum sciences. The ability to produce entangled particles in a controllable manner is essential for any quantum technology. Entanglement between light particles (photons) is particularly crucial for quantum communication due to light's non-interactive nature and long-lasting coherence. Resources producing entangled multiphoton cluster states will enable communication between remote quantum nodes, as the inbuilt redundancy of cluster photons allows for repeated local measurements-compensating for losses and probabilistic Bell measurements. For feasible applications, the cluster generation should be fast, deterministic and, most importantly, its photons indistinguishable, which will allow measurements and fusion of clusters by interfering photons. Here, using periodic excitation of a semiconductor quantum-dot-confined spin, we demonstrate a multi-indistinguishable photon cluster, featuring a continuously generated string of photons at deterministic gigahertz generation rates, and an optimized entanglement length of about ten photons. The indistinguishability of the photons opens up new possibilities for scaling up the cluster's dimensionality by fusion, thus building graph states suited for measurement-based photonic quantum computers and all-photonic quantum repeaters.

Interactions between out-of-plane dipoles in bosonic gases enable the long-range propagation of excitons. The lack of direct control over collective dipolar properties has so far limited the degrees of tunability and the microscopic understanding of exciton transport. In this work we modulate the layer hybridization and interplay between many-body interactions of excitons in a van der Waals heterostructure with an applied vertical electric field. By performing spatiotemporally resolved measurements supported by microscopic theory, we uncover the dipole-dependent properties and transport of excitons with different degrees of hybridization. Moreover, we find constant emission quantum yields of the transporting species as a function of excitation power with radiative decay mechanisms dominating over nonradiative ones, a fundamental requirement for efficient excitonic devices. Our findings provide a complete picture of the many-body effects in the transport of dilute exciton gases, and have crucial implications for studying emerging states of matter such as Bose-Einstein condensation and optoelectronic applications based on exciton propagation.

Quantum key distribution has emerged as the most viable scheme to guarantee information security in the presence of large-scale quantum computers and, thanks to the continuous progress made in the past 20 years, it is now commercially available. However, the secret key rates remain limited to just over 10 Mbps due to several bottlenecks on the receiver side. Here we present a custom multipixel superconducting nanowire single-photon detector that is designed to guarantee high count rates and precise timing discrimination. Leveraging the performance of the detector and coupling it to fast acquisition and real-time key distillation electronics, we remove two major roadblocks and achieve a considerable increase of the secret key rates with respect to the state of the art. In combination with a simple 2.5-GHz clocked time-bin quantum key distribution system, we can generate secret keys at a rate of 64 Mbps over a distance of 10.0 km and at a rate of 3.0 Mbps over a distance of 102.4 km with real-time key distillation.

A fundamental question regarding light scattering is how whiteness, generated from multiple scattering, can be obtained from thin layers of materials. This challenge arises from the phenomenon of optical crowding, whereby, for scatterers packed with filling fractions higher than ~30%, reflectance is drastically reduced due to near-field coupling between the scatterers. Here we show that the extreme birefringence of isoxanthopterin nanospheres overcomes optical crowding effects, enabling multiple scattering and brilliant whiteness from ultra-thin chromatophore cells in shrimp. Strikingly, numerical simulations reveal that birefringence, originating from the spherulitic arrangement of isoxanthopterin molecules, enables intense broadband scattering almost up to the maximal packing for random spheres. This reduces the thickness of material required to produce brilliant whiteness, resulting in a photonic system that is more efficient than other biogenic or biomimetic white materials which operate in the lower refractive index medium of air. These results highlight the importance of birefringence as a structural variable to enhance the performance of such materials and could contribute to the design of biologically inspired replacements for artificial scatterers like titanium dioxide.

Deep neural networks have achieved remarkable breakthroughs by leveraging multiple layers of data processing to extract hidden representations, albeit at the cost of large electronic computing power. To enhance energy efficiency and speed, the optical implementation of neural networks aims to harness the advantages of optical bandwidth and the energy efficiency of optical interconnections. In the absence of low-power optical nonlinearities, the challenge in the implementation of multilayer optical networks lies in realizing multiple optical layers without resorting to electronic components. Here we present a novel framework that uses multiple scattering, and which is capable of synthesizing programmable linear and nonlinear transformations concurrently at low optical power by leveraging the nonlinear relationship between the scattering potential, represented by data, and the scattered field. Theoretical and experimental investigations show that repeating the data by multiple scattering enables nonlinear optical computing with low-power continuous-wave light. Moreover, we empirically find that scaling of this optical framework follows a power law.