Ultrasensitive Vibrational Imaging of Retinoids by Visible Preresonance Stimulated Raman Scattering Microscopy

Published on Feb 8, 2021in Advanced Science15.84
· DOI :10.1002/ADVS.202003136
Minghua Zhuge1
Estimated H-index: 1
(ZJU: Zhejiang University),
Kai-Chih Huang9
Estimated H-index: 9
(BU: Boston University)
+ 8 AuthorsJi-Xin Cheng83
Estimated H-index: 83
(BU: Boston University)
High-sensitivity chemical imaging offers a window to decipher the molecular orchestra inside a living system. Based on vibrational fingerprint signatures, coherent Raman scattering microscopy provides a label-free approach to map biomolecules and drug molecules inside a cell. Yet, by near-infrared (NIR) pulse excitation, the sensitivity is limited to millimolar concentration for endogenous biomolecules. Here, the imaging sensitivity of stimulated Raman scattering (SRS) is significantly boosted for retinoid molecules to 34 micromolar via electronic preresonance in the visible wavelength regime. Retinoids play critical roles in development, immunity, stem cell differentiation, and lipid metabolism. By visible preresonance SRS (VP-SRS) imaging, retinoid distribution in single embryonic neurons and mouse brain tissues is mapped, retinoid storage in chemoresistant pancreatic and ovarian cancers is revealed, and retinoids stored in protein network and lipid droplets of Caenorahbditis elegans are identified. These results demonstrate VP-SRS microscopy as an ultrasensitive label-free chemical imaging tool and collectively open new opportunities of understanding the function of retinoids in biological systems.
#1Kai-Chih Huang (BU: Boston University)H-Index: 9
#2Junjie Li (BU: Boston University)H-Index: 16
Last. Ji-Xin Cheng (BU: Boston University)H-Index: 83
view all 5 authors...
Summary In situ measurement of cellular metabolites is still a challenge in biology. Conventional methods, such as mass spectrometry or fluorescence microscopy, would either destruct the sample or introduce strong perturbations to target molecules. Here, we present multiplex stimulated Raman scattering (SRS) imaging cytometry as a label-free single-cell analysis platform with chemical specificity and high-throughput capabilities. Using SRS imaging cytometry, we studied the metabolic responses of...
7 CitationsSource
#1Hyeon Jeong Lee (BU: Boston University)H-Index: 10
#2Kai-Chih Huang (BU: Boston University)H-Index: 9
Last. Ji-Xin ChengH-Index: 83
view all 8 authors...
Voltage imaging allows mapping of membrane potential in living cells. Yet, current intensity-based imaging approaches are limited to relative membrane potential changes, missing important information conveyed by the absolute value of membrane voltage. This challenge arises from various factors affecting the signal intensity, such as concentration, illumination intensity, and photobleaching. Here, we demonstrate electronic pre-resonance hyperspectral stimulated Raman scattering (EPR-hSRS) for spe...
2 CitationsSource
#1Luyuan Zhang (Columbia University)H-Index: 9
#2Lingyan Shi (Columbia University)H-Index: 14
Last. Wei Min (Columbia University)H-Index: 39
view all 8 authors...
Cells and tissues often display pronounced spatial and dynamical metabolic heterogeneity. Common glucose-imaging techniques report glucose uptake or catabolism activity, yet do not trace the functional utilization of glucose-derived anabolic products. Here we report a microscopy technique for the optical imaging, via the spectral tracing of deuterium (STRIDE), of diverse macromolecules derived from glucose. Based on stimulated Raman-scattering imaging, STRIDE visualizes the metabolic dynamics of...
25 CitationsSource
#1Minbiao Ji (Fudan University)H-Index: 23
#2Michal Arbel (Harvard University)H-Index: 2
Last. X. Sunney Xie (Harvard University)H-Index: 100
view all 9 authors...
One of the key pathological features of Alzheimer’s disease (AD) is the existence of extracellular deposition of amyloid plaques formed with misfolded amyloid-β (Aβ). The conformational change of proteins leads to enriched contents of β sheets, resulting in remarkable changes of vibrational spectra, especially the spectral shifts of the amide I mode. Here, we applied stimulated Raman scattering (SRS) microscopy to image amyloid plaques in the brain tissue of an AD mouse model. We have demonstrat...
40 CitationsSource
#1Yali Bi (CCNU: Central China Normal University)H-Index: 2
#2Chi Yang (HUST: Huazhong University of Science and Technology)H-Index: 1
Last. Ping Wang (HUST: Huazhong University of Science and Technology)H-Index: 17
view all 8 authors...
High-resolution optical microscopes that can break 180 nm in spatial resolution set to conventional microscopies are much-needed tools. However, current optical microscopes have to rely on exogenous fluorescent labels to achieve high resolution in biological imaging. Herein, we report near-resonance enhanced label-free stimulated Raman scattering (SRS) microscopy with a lateral resolution near 130 nm, in which the high-resolution image contrast originates directly from a low concentration of end...
25 CitationsSource
#1Andy J. Chen (Purdue University)H-Index: 3
#1Andy Chen (Purdue University)H-Index: 10
Last. Ji-Xin Cheng (BU: Boston University)H-Index: 83
view all 6 authors...
: Retinoids play critical roles in development, immunity, and lipid metabolism, and their deficiency leads to various human disorders. Yet, tools for sensing retinoids in vivo are lacking, which limits the understanding of retinoid distribution, dynamics and functions in living organisms. Here, using hyperspectral stimulated Raman scattering microscopy, we discover a previously unknown cytoplasmic store of retinoids in Caenorahbditis elegans. Following the temporal dynamics of retinoids, we reve...
6 CitationsSource
#1Lixue Shi (Columbia University)H-Index: 6
#2Hanqing Xiong (Columbia University)H-Index: 7
Last. Wei Min (Columbia University)H-Index: 39
view all 6 authors...
Recently we have reported electronic pre-resonance stimulated Raman scattering (epr-SRS) microscopy as a powerful technique for super-multiplex imaging (Wei, L.; Nature 2017, 544, 465−470). However, under rigorous electronic resonance, background signal, which mainly originates from pump–probe process, overwhelms the desired vibrational signature of the chromophores. Here we demonstrate electronic resonant stimulated Raman scattering (er-SRS) microspectroscopy and imaging through suppression of ...
14 CitationsSource
#1Michael M. Gaschler (Columbia University)H-Index: 7
#2Fanghao Hu (Columbia University)H-Index: 22
Last. Brent R. Stockwell (Columbia University)H-Index: 76
view all 6 authors...
Ferroptosis is a form of nonapoptotic cell death characterized by the unchecked accumulation of lipid peroxides. Ferrostatin-1 and its analogs (ferrostatins) specifically prevent ferroptosis in multiple contexts, but many aspects of their molecular mechanism of action remain poorly described. Here, we employed stimulated Raman scattering (SRS) microscopy coupled with small vibrational tags to image the distribution of ferrostatins in cells and found that they accumulate in lysosomes, mitochondri...
80 CitationsSource
#1Yihui Shen (Columbia University)H-Index: 18
#2Zhilun Zhao (Columbia University)H-Index: 6
Last. Wei Min (Columbia University)H-Index: 39
view all 8 authors...
Membrane phase behavior has been well characterized in model membranes in vitro under thermodynamic equilibrium state. However, the widely observed differences between biological membranes and their in vitro counterparts are placing more emphasis on nonequilibrium factors, including influx and efflux of lipid molecules. The endoplasmic reticulum (ER) is the largest cellular membrane system and also the most metabolically active organelle responsible for lipid synthesis. However, how the nonequil...
41 CitationsSource
#1Lu Wei (Columbia University)H-Index: 19
#2Zhixing Chen (Columbia University)H-Index: 18
Last. Wei Min (Columbia University)H-Index: 39
view all 10 authors...
The ability to visualize directly a large number of distinct molecular species inside cells is increasingly essential for understanding complex systems and processes. Even though existing methods have successfully been used to explore structure–function relationships in nervous systems, to profile RNA in situ, to reveal the heterogeneity of tumour microenvironments and to study dynamic macromolecular assembly, it remains challenging to image many species with high selectivity and sensitivity und...
137 CitationsSource
Cited By1
#1Jiabao Xu (University of Oxford)
#1Jiabao XuH-Index: 8
Last. Wei E. HuangH-Index: 40
view all 7 authors...
Metabolic reprogramming is a common hallmark in cancer. The high complexity and heterogeneity in cancer render it challenging for scientists to study cancer metabolism. Despite the recent advances in single-cell metabolomics based on mass spectrometry, the analysis of metabolites is still a destructive process, thus limiting in vivo investigations. Being label-free and nonperturbative, Raman spectroscopy offers intrinsic information for elucidating active biochemical processes at subcellular lev...