arXiv subjects

Guangrui Ding

Publications and source records attributed to Guangrui Ding.

7 recordsLinked to original sources

arXiv2026

FILM: Mapping organellar metabolism by mid-infrared photothermal modulated fluorescence

Metabolism unfolds within specific organelles in eukaryotic cells. Lysosomes are highly metabolically active organelles, and their metabolic states dynamically influence signal transduction, cellular homeostasis, and organismal physiopathology. Despite the significance of lysosomal metabolism, a method for its in vivo measurement is currently lacking. Here, we report optical boxcar-enhanced, fluorescence-detected mid-infrared photothermal microscopy, together with AI-assisted data denoising and spectral deconvolution, to map metabolic activity and composition of individual lysosomes in living cells and organisms. Using this method, we uncovered lipolysis and proteolysis heterogeneity across lysosomes within the same cell, as well as early-onset lysosomal dysfunction during organismal aging. Additionally, we discovered organelle-level metabolic changes associated with diverse lysosomal storage diseases. This method holds the broad potential to profile metabolic fingerprints of individual organelles within their native context and quantitatively assess their dynamic changes under different physiological and pathological conditions, providing a high-resolution chemical cellular atlas.

physics.bio-ph↗

arXiv2026

Label-free Imaging of Single-Biomolecule Structure and Interaction by Stimulated Raman Photothermal Encoded Scattering

Current single molecule methods either rely on fluorescence or lack chemical information. Here we report stimulated Raman photothermal encoded scattering (SRPSCAT) microscopy for quantitative bond-selective imaging of single-biomolecule structures and interactions in native environments. In this approach, scattering of the target molecule is modulated by the deposited energy from stimulated Raman gain and loss processes, thereby encoding vibrational spectroscopic information. Leveraging single-molecule sensitivity of interferometric scattering, SRPSCAT can map single proteins with chemical specificity, determine their mass, and distinguish protein secondary structures based on their Raman fingerprints. Furthermore, single protein binding kinetics are quantified and the conformational dynamics of single de novo designed allosteric proteins are observed. Together, these results highlight the potential of SRPSCAT for label-free structural, functional and dynamic analysis at the single-molecule level.

physics.bio-ph↗

arXiv2026

Mid-wave infrared photothermal microscopy for molecular and metabolic imaging in deep tissues and spheroids

High-resolution chemical imaging within deep tissues and intact spheroids remains a grand challenge. Here, we introduce mid-wave infrared photothermal (MWIP) microscopy operating in the underexplored 2000-2500 nm spectral window for submicron-resolution molecular and metabolic imaging in intact tumor spheroids and deep tissues. A dark-field photothermal detection scheme significantly suppresses water background and enhances contrast. By accessing strong carbon-hydrogen combination absorptions, a detection limit of 0.12% for dimethyl sulfoxide is achieved, comparable to stimulated Raman scattering microscopy. Depth-resolved imaging of endogenous biomolecules up to 500 micrometers in excised mouse skin and brain tissues is demonstrated. MWIP further enables depth-resolved tracking of transdermal drug transport via carbon-deuterium overtone absorption. Using deuterium metabolic probes, fatty-acid metabolism is imaged at 200 micrometers deep within intact tumor spheroids through carbon-deuterium overtone and combination bands. Collectively, MWIP offers a platform for functional imaging of 3D biological systems in their native environments.

physics.optics↗

arXiv2026

Micromolar chemical imaging by high-energy low-photodamage Coherent Anti-stokes Raman Scattering (HELP-CARS)

Coherent anti-Stokes Raman scattering (CARS) microscopy offers label-free chemical imaging capabilities, but its performance is constrained by small Raman scattering cross-section, strong non-resonant background (NRB), and limited signal-to-noise ratio (SNR). Here, we introduce a high-energy, low-photodamage CARS (HELP-CARS) platform designed to overcome these physical limitations. By employing a 1-MHz non-collinear optical parametric amplifier (NOPA) with extensive pulse chirping, HELP-CARS increases the coherent Raman excitation efficiency by ~300 times and improves the signal-to-nonresonant background ratio by 11 times, while inducing negligible damage during live cell imaging. Furthermore, to remove non-independent noise and physically entangled non-resonant background, we incorporate self-supervised deep-learning denoising and background removal based on the Kramers-Kronig relationship, yielding sensitivity improvement by an additional order of magnitude. Together, these advances provide a micromolar imaging sensitivity (160 uM for Dimethyl sulfoxide-d6) corresponding to 1000 molecules in the focal volume. Such high sensitivity enables high-fidelity chemical imaging in both fingerprint and silent windows. Hyperspectral HELP-CARS imaging of deuterated fatty acids allowed first observation of chemical separation with single lipid droplet. Together, HELP-CARS offers a powerful and generalizable approach for ultrasensitive and quantitative vibrational imaging of biological systems.

physics.optics↗

arXiv2025

Spatial-offset pump-probe imaging of nonradiative dynamics at optical resolution

Nonradiative photothermal (PT) and photoacoustic (PA) processes have found widespread applications in imaging, stimulation, and therapy. Mapping the generation and propagation of PA and PT waves with resolution is important to elucidate how these fields interact with biological systems. To this end, we introduce spatial offset pump-probe imaging (SOPPI). By spatially offsetting the pump beam and the probe beam, SOPPI can image simultaneously PA and PT wave propagation with nanosecond temporal resolution, micrometer spatial resolution, 65 MHz detection bandwidth, and a sensitivity of 9.9 Pa noise equivalent pressure. We first map the PA and PT evolution from a fiber emitter, and how the wave interacting with a mouse skull and brain slices. SOPPI imaging of PA waves from a tapered fiber with water as an absorber shows a wavelength-dependent generation, evanescent wave generated PA, and back-propagated acoustic Mach Cone. At last, a SOPPI-PACT is developed to reconstruct the pigment distribution inside a zebrafish larva with high precision and signal-to-noise ratio.

physics.optics↗

arXiv2024

Self-Supervised Elimination of Non-Independent Noise in Hyperspectral Imaging

Hyperspectral imaging has been widely used for spectral and spatial identification of target molecules, yet often contaminated by sophisticated noise. Current denoising methods generally rely on independent and identically distributed noise statistics, showing corrupted performance for non-independent noise removal. Here, we demonstrate Self-supervised PErmutation Noise2noise Denoising (SPEND), a deep learning denoising architecture tailor-made for removing non-independent noise from a single hyperspectral image stack. We utilize hyperspectral stimulated Raman scattering and mid-infrared photothermal microscopy as the testbeds, where the noise is spatially correlated and spectrally varied. Based on single hyperspectral images, SPEND permutates odd and even spectral frames to generate two stacks with identical noise properties, and uses the pairs for efficient self-supervised noise-to-noise training. SPEND achieved an 8-fold signal-to-noise improvement without having access to the ground truth data. SPEND enabled accurate mapping of low concentration biomolecules in both fingerprint and silent regions, demonstrating its robustness in sophisticated cellular environments.

eess.IV↗

arXiv2020

Generalized Perfect Optical Vortex along Arbitrary Trajectories

Perfect optical vortex (POV) is a type of vortex beam with an infinite thin ring and a fixed radius independent of its topological charge. Here we propose the concept of generalized perfect optical vortex along arbitrary curves beyond the regular shapes of circle and ellipse. Generalized perfect optical vortices also share the similar properties as POVs, such as defined only along infinite thin curves and owning topological charges independent of scales. Notably, they naturally degenerate to the POVs and elliptic POVs along circles and ellipses, respectively. We also experimentally generated the generalized perfect optical vortices through a digital micromirror device (DMD) and measured the phase distributions by interferometry, exhibiting good agreements with the simulations. Moreover, we derive a proper modified formula to yield the generalized perfect optical vortices with uniform intensity distribution along predesigned curves. The generalized perfect optical vortices might find the potential applications in optical tweezers and communication.

physics.optics↗