State-of-the-art microscopes use intense lasers that can severely disturb biological processes, function and viability. This introduces hard limits on performance that only quantum photon correlations can overcome. Here we demonstrate this absolute quantum advantage, achieving signal-to-noise beyond the photodamage-free capacity of conventional microscopy. We achieve this in a coherent Raman microscope, which we use to image molecular bonds within a cell with both quantum-enhanced contrast and sub-wavelength resolution. This allows the observation of nanoscale biological structures that would otherwise not be resolved. Coherent Raman microscopes allow highly selective biomolecular finger-printing in unlabelled specimens, but photodamage is a major roadblock for many applications. By showing that this roadblock can be overcome, our work provides a path towards order-of-magnitude improvements in both sensitivity and imaging speed.
Label-free nonlinear optical microscopy has become a powerful tool for biomedical research. However, the possible photodamage risk hinder further clinical applications. To reduce these adverse effects, we constructed a new platform of simultaneous label-free autofluorescence multiharmonic (SLAM) microscopy, featuring 5-channel multimodal imaging, inline photodamage monitoring, and pulse repetition-rate tuning. By the use of a birefringent photonic crystal fiber for spectral broadening (rather than supercontinuum generation) and a prism compressor for pulse pre-chirping, this system allows user…
Micromolar chemical imaging by high-energy low-photodamage Coherent Anti-stokes Raman Scattering (HELP-CARS)
Boston University（波士顿大学）；The Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University（佐治亚理工学院和埃默里大学华莱士·H·考尔特生物医学工程系）
Two-photon light-sheet live imaging at kilohertz frame rate using birefringence-based pulse splitting
Laboratoire d’Optique et Biosciences, CNRS, INSERM, École Polytechnique, Institut Polytechnique de Paris（光学与生物科学实验室，法国国家科学研究中心，法国国家健康与医学研究院，巴黎综合理工学院，巴黎理工学院）；ICube Research Institute, CNRS, University of Strasbourg（ICube研究所，法国国家科学研究中心，斯特拉斯堡大学）；Sorbonne Université, CNRS, Laboratoire Jean Perrin（索邦大学，法国国家科学研究中心，让·佩兰实验室）
Conventional linear optical activity effects are widely used for studying chiral materials. However, poor contrast and artifacts due to sample anisotropy limit the applicability of these methods. Here we demonstrate that nonlinear second-harmonic-generation circular dichroism spectral microscopy can overcome these limits. In intact collagenous tissues, clear spectral resonance is observed with sub-micrometer spatial resolution. By performing gradual protein denaturation studies, we show that the resonant responses are dominantly due to the molecular chirality.
Optical microscopy has so far been restricted to superficial layers, leaving many important biological questions unanswered. Random scattering causes the ballistic focus, which is conventionally used for image formation, to decay exponentially with depth. Optical imaging beyond the ballistic regime has been demonstrated by hybrid techniques that combine light with the deeper penetration capability of sound waves. Deep inside highly scattering media, the sound focus dimensions restrict the imaging resolutions. Here we show that by iteratively focusing light into an ultrasound focus via phase co…
In recent years several far-field microscopy techniques have been developed which manage to overcome the diffraction limit of resolution. A unifying classification scheme for them is clearly desirable. We argue that existing schemes based on the information capacity of the optical system can not easily be extended to cover e.g., STED microscopy or techniques based on single molecule imaging. We suggest a classification based on a reconstruction of the Abbe limit.
We demonstrate a new 'microsphere nanoscope' that uses ordinary SiO2 microspheres as superlenses to create a virtual image of the object in near field. The magnified virtual image greatly overcomes the diffraction limit. We are able to resolve clearly 50-nm objects under a standard white light source in both transmission and reflection modes. The resolution achieved for white light opens a new opportunity to image viruses, DNA and molecules in real time.
Label-free imaging of rapidly moving, sub-diffraction sized structures has important applications in both biology and material science, as it removes the limitations associated with fluorescence tagging. However, unlabeled nanoscale particles in suspension are difficult to image due to their transparency and fast Brownian motion. Here we describe a novel interferometric imaging technique referred to as Magnified Image Spatial Spectrum (MISS) microscopy, which overcomes these challenges. The MISS microscope provides quantitative phase information and enables dynamic light scattering investigati…
Raman sensing and Raman microscopy are amongst the most specific optical technologies to identify the chemical compounds of unknown samples, and to enable label-free biomedical imaging with molecular contrast. However, the high cost and complexity, low speed, and incomplete spectral information provided by current technology are major challenges preventing more widespread application of Raman systems. To overcome these limitations, we developed a new method for stimulated Raman spectroscopy and Raman imaging using continuous wave (CW), rapidly wavelength swept lasers. Our all-fiber, time-encod…
In recent years several methods to overcome diffraction limit in the far field microscopy have been demonstrated. Still the problem of superresolution is reliably solved only for fluorescent microscopy, giving a resolution of up to 20-30nm. Obtaining the optical resolution lower than 100nm without fluorescent dyes requires using rather slow and complicated technique of scanning near filed optical microscope (SNOM). We propose and demonstrate a method of optical near field acquisition by far-field microscope through observation of nanoparticles Brownian motion in immersion liquid. The resolutio…
High space-bandwidth product with high spatial phase sensitivity is indispensable for a single-shot quantitative phase microscopy (QPM) system. It opens avenue for widespread applications of QPM in the field of biomedical imaging. Temporally low coherence length light sources are generally implemented to achieve high spatial phase sensitivity in QPM at the cost of either reduced temporal resolution or smaller field of view (FOV). On the contrary, high temporal coherence light sources like lasers are capable of exploiting the full FOV of the QPM systems at the expense of less spatial phase sens…
We propose and demonstrate a photon-efficient optical classifier to overcome the Rayleigh limit in spatial resolution. It utilizes mode-selective sum frequency generation and single-pixel photon detection to resolve closely spaced incoherent sources based on photon counting statistics. Super-resolving and photon efficient, this technique can find applications in microscopy, light detection and ranging (LiDAR), and astrophysics.
Super-resolution light microscopy overcomes the physical barriers due to light diffraction, allowing for the observation of otherwise indistinguishable subcellular entities. However, the specific acquisition conditions required by state-of-the-art super-resolution methods to achieve adequate spatio-temporal resolution are often very challenging. Exploiting molecules fluctuations allows good spatio-temporal resolution live-cell imaging by means of common microscopes and conventional fluorescent dyes. In this work, we present the method COL0RME for COvariance-based $\ell_0$ super-Resolution Micr…
Brillouin microscopy has recently emerged as powerful technique to characterize the mechanical properties of biological tissue, cell and biomaterials. However, the potential of Brillouin microscopy is currently limited to transparent samples, because Brillouin spectrometers do not have sufficient spectral extinction to reject the predominant non-Brillouin scattered light of turbid media. To overcome this issue, we developed a spectrometer composed of a two VIPA stages and a multi-pass Fabry-Perot interferometer. The Fabry-Perot etalon acts as an ultra-narrow band-pass filter for Brillouin ligh…
Super-resolution fluorescence microscopy overcomes blurring arising from light diffraction, allowing the reconstruction of fine scale details in biological structures. Standard methods come at the expense of long acquisition time and/or harmful effects on the biological sample, which makes the problem quite challenging for the imaging of body cells. A promising new avenue is the exploitation of molecules fluctuations, allowing live-cell imaging with good spatio-temporal resolution through common microscopes and conventional fluorescent dyes. Several numerical algorithms have been developed in …
Quantum-optical coherence tomography (QOCT) is an optical sectioning modality based on the quantum interference of photon pairs, obtained from a spontaneous parametric downconversion (SPDC) source. The promise of QOCT derives from two quantum-conferred advantages when compared to equivalent classical optical coherence tomography (OCT) systems: a factor of 2 axial resolution enhancement, as well as dispersion cancellation. However, the technique is far from being competitive with current OCT devices due to the long required acquisition times. Here, we on the one hand demonstrate a quantum optic…
Fourier ptychographic microscopy (FPM) is a computational imaging technique that overcomes the physical space-bandwidth product (SBP) limit of a conventional microscope by applying angular diversity illuminations. In the usual model of FPM, the microscopic system is approximated by being taken as space-invariant with transfer function determined by a complex pupil function of the objective. However, in real experimental conditions, several unexpected "semi-bright and semi-dark" images with strong vignetting effect can be easily observed when the sample is illuminated by the LED within the "tra…
This work presents the development of a flexible microscopic chemical imaging platform for cultural heritage that utilizes wavelength-tunable oblique illumination from a point source to obtain per-pixel reflectance spectra in the VIS-NIR range. The microscope light source can be adjusted on two axes allowing for a hemisphere of possible illumination directions. The synthesis of multiple illumination angles allows for the calculation of surface normal vectors, similar to phase gradients, and axial optical sectioning. The extraction of spectral reflectance images with high spatial resolutions fr…
By using scattering in near field techniques, a microscope can be easily turned into a device measuring static and dynamic light scattering, very useful for the characterization of nanoparticle dispersions. Up to now, microscopy based techniques have been limited to forward scattering, up to a maximum of 30 degrees. In this paper we present a novel optical scheme that overcomes this limitation, extending the detection range to angles larger than 90 degrees (back-scattering). Our optical scheme is based on a microscope, a wide numerical aperture objective, and a laser illumination, with the col…
The diffraction limit is a fundamental barrier in optical microscopy, which restricts the smallest resolvable feature size of a microscopic system. Microsphere-based microscopy has proven to be a promosing tool for challenging the diffraction limit. Nevertheless, the microspheres have a low imaging contrast in the air, which hinders the application of this technique. In this Letter, we demonstrate that this challenge can be effectively overcome by using partially Ag-plated microspheres. The deposited Ag film acts as an aperture stop that blocks a portion of the incident beam, forming a photoni…
Institute of Micro- and Nanotechnologies, Technische Universität Ilmenau（微纳技术研究所，埃尔朗根-纽伦堡工业大学）；Institute of Thermodynamics and Fluid Mechanics, Technische Universität Ilmenau（热力学与流体力学研究所，埃尔朗根-纽伦堡工业大学）
Pic2Spec: Generative Modeling Reconstructs Single Cell Raman Fingerprints from Brightfield Images
Department of Mechanical Engineering, MIT（麻省理工学院机械工程系）；Ragon Institute of Massachusetts General Hospital, MIT, and Harvard（麻省总医院、麻省理工学院和哈佛大学的拉根研究所）；Jameel Clinic for AI & Healthcare, MIT（麻省理工学院贾米尔人工智能与医疗诊所）
