Biological imaging of chemical bonds by stimulated Raman scattering microscopy

Nature Methods, Journal Year: 2019, Volume and Issue: 16(9), P. 830 - 842

Published: Aug. 30, 2019

Language: Английский

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Phenazine production promotes antibiotic tolerance and metabolic heterogeneity in Pseudomonas aeruginosa biofilms

Nature Communications, Journal Year: 2019, Volume and Issue: 10(1)

Published: Feb. 15, 2019

Abstract Antibiotic efficacy can be antagonized by bioactive metabolites and other drugs present at infection sites. Pseudomonas aeruginosa , a common cause of biofilm-based infections, releases called phenazines that accept electrons to support cellular redox balancing. Here, we find promote tolerance clinically relevant antibiotics, such as ciprofloxacin, in P. biofilms this effect depends on the carbon source provided for growth. We couple stable isotope labeling with stimulated Raman scattering microscopy visualize biofilm metabolic activity situ. This approach shows metabolism microaerobic regions influence responses ciprofloxacin treatment. Consistent roles specific respiratory complexes supporting phenazine utilization biofilms, phenazine-dependent survival is diminished mutants lacking these enzymes. Our work introduces technique chemical imaging biosynthetic highlights complex interactions between bacterial products, their effects metabolism, antibiotics use treat infections.

Language: Английский

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Raman Spectroscopy for Chemical Biology Research

Journal of the American Chemical Society, Journal Year: 2022, Volume and Issue: 144(43), P. 19651 - 19667

Published: Oct. 10, 2022

In chemical biology research, various fluorescent probes have been developed and used to visualize target proteins or molecules in living cells tissues, yet there are limitations this technology, such as the limited number of colors that can be detected simultaneously. Recently, Raman spectroscopy has applied overcome limitations. detects molecular vibrations reflecting structures conditions a sample was originally directly responses endogenous molecules. However, our initial research develop "Raman tags" opens new avenue for application biology. Perspective, we first introduce label-free imaging biomolecules, illustrating biological applications spectroscopy. Next, highlight small using tags research. Finally, discuss development potential probes, which represent next-generation

Language: Английский

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Protein and lipid mass concentration measurement in tissues by stimulated Raman scattering microscopy

Proceedings of the National Academy of Sciences, Journal Year: 2022, Volume and Issue: 119(17)

Published: April 22, 2022

Significance We report a quantitative Raman microscopy method that measures the concentration of protein and lipid in cells at high spatial resolution living fixed samples tissues, allowing studies cell size organelle regulation both culture tissue slices; it can be applied to problems control, intracellular crowding, metabolism context growth, differentiation, senescence, pathology.

Language: Английский

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Super-resolution SRS microscopy with A-PoD

Nature Methods, Journal Year: 2023, Volume and Issue: 20(3), P. 448 - 458

Published: Feb. 16, 2023

Language: Английский

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Microglial lipid droplet accumulation in tauopathy brain is regulated by neuronal AMPK

Cell Metabolism, Journal Year: 2024, Volume and Issue: 36(6), P. 1351 - 1370.e8

Published: April 23, 2024

Language: Английский

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Single-cell mapping of lipid metabolites using an infrared probe in human-derived model systems

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: Jan. 8, 2024

Abstract Understanding metabolic heterogeneity is the key to uncovering underlying mechanisms of metabolic-related diseases. Current imaging studies suffer from limitations including low resolution and specificity, model systems utilized often lack human relevance. Here, we present a single-cell platform enable direct lipid metabolism with high specificity in various human-derived 2D 3D culture systems. Through incorporation an azide-tagged infrared probe, selective detection newly synthesized lipids cells tissue became possible, while simultaneous fluorescence enabled cell-type identification complex tissues. In proof-of-concept experiments, were directly visualized human-relevant among different cell types, mutation status, differentiation stages, over time. We identified upregulated progranulin-knockdown induced pluripotent stem their differentiated microglia cells. Furthermore, observed that neurons brain organoids exhibited significantly lower compared astrocytes.

Language: Английский

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Multi-molecular hyperspectral PRM-SRS microscopy

Nature Communications, Journal Year: 2024, Volume and Issue: 15(1)

Published: Feb. 21, 2024

Abstract Lipids play crucial roles in many biological processes. Mapping spatial distributions and examining the metabolic dynamics of different lipid subtypes cells tissues are critical to better understanding their aging diseases. Commonly used imaging methods (such as mass spectrometry-based, fluorescence labeling, conventional optical imaging) can disrupt native environment cells/tissues, have limited or spectral resolution, cannot distinguish subtypes. Here we present a hyperspectral platform that integrates Penalized Reference Matching algorithm with Stimulated Raman Scattering (PRM-SRS) microscopy. Using this platform, visualize identify high density lipoprotein particles human kidney, cholesterol phosphatidylethanolamine ratio inside granule mouse hippocampus, subcellular sphingosine cardiolipin brain. Our PRM-SRS displays unique advantages enhanced chemical specificity, fast data processing distinguishing organs species.

Language: Английский

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Rapid Antibiotic Susceptibility Testing of Pathogenic Bacteria Using Heavy-Water-Labeled Single-Cell Raman Spectroscopy in Clinical Samples

Analytical Chemistry, Journal Year: 2019, Volume and Issue: 91(9), P. 6296 - 6303

Published: April 3, 2019

Speeding up antibiotic susceptibility testing (AST) is urgently needed in clincial settings to guide fast and tailored prescription before treatment. It remains a big challenge achieve sample-to-AST answer within half working day directly from clinical sample. Here we develop single-cell Raman spectroscopy coupled with heavy water labeling (Raman-D2O) as rapid activity-based AST approach applicable for urine samples. By rapidly transferring (15 min) bacteria AST, the total assay time receiving binary susceptibility/resistance (S/R) readout was shortened only 2.5 h. Moreover, by overcoming nonsynchronous responses between microbial activity growth, together setting new S/R cutoff value based on relative C-D ratios, of both pathogenic isolates three urines against antibiotics different action mechanisms determined Raman-D2O were all consistent slow standard used settings. This work promotes practicability faciliates stewardship.

Language: Английский

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Volumetric chemical imaging by clearing-enhanced stimulated Raman scattering microscopy

Proceedings of the National Academy of Sciences, Journal Year: 2019, Volume and Issue: 116(14), P. 6608 - 6617

Published: March 14, 2019

Significance Cells form structures and perform functions through intricate 3D tissue organizations. However, due to scattering, coherent Raman microscopy—a powerful method complementary fluorescence imaging—suffers from limited imaging depth in tissues. Here, we develop a volumetric chemical with greater than 10-fold increase. We formulate Raman-tailored tissue-clearing recipe combine it advanced microscopies. Equipped the toolbox of analyses, elucidate complex structures, compositions, metabolic dynamics diverse tissues including lipid synthesis throughout tumor spheroids, networks axons, vasculatures, cell bodies brain regions, as well heterogeneous metabolism.

Language: Английский

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