Multicolour chemical imaging of plant tissues with hyperspectral stimulated Raman scattering microscopy

The Analyst ◽  
2021 ◽  
Author(s):  
Takanori Iino ◽  
Kenji Hashimoto ◽  
Takuya Asai ◽  
Kazuyuki Kuchitsu ◽  
Yasuyuki Ozeki
Keyword(s):  
Raman Scattering ◽  
Stimulated Raman Scattering ◽  
Chemical Imaging ◽  
Plant Tissues ◽  
Label Free ◽  
Stimulated Raman

Stimulated Raman scattering microscopy allows for multicolour label-free chemical imaging of plant tissues.

scholarly journals Label-free chemical imaging flow cytometry by high-speed multicolor stimulated Raman scattering

2019 ◽  
Vol 116 (32) ◽  
pp. 15842-15848 ◽  
Author(s):  
Yuta Suzuki ◽  
Koya Kobayashi ◽  
Yoshifumi Wakisaka ◽  
Dinghuan Deng ◽  
Shunji Tanaka ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Raman Scattering ◽  
Cancer Biology ◽  
High Speed ◽  
Stimulated Raman Scattering ◽  
Chemical Imaging ◽  
Fluorescent Labeling ◽  
Label Free ◽  
Imaging Flow Cytometry ◽  
Stimulated Raman

Combining the strength of flow cytometry with fluorescence imaging and digital image analysis, imaging flow cytometry is a powerful tool in diverse fields including cancer biology, immunology, drug discovery, microbiology, and metabolic engineering. It enables measurements and statistical analyses of chemical, structural, and morphological phenotypes of numerous living cells to provide systematic insights into biological processes. However, its utility is constrained by its requirement of fluorescent labeling for phenotyping. Here we present label-free chemical imaging flow cytometry to overcome the issue. It builds on a pulse pair-resolved wavelength-switchable Stokes laser for the fastest-to-date multicolor stimulated Raman scattering (SRS) microscopy of fast-flowing cells on a 3D acoustic focusing microfluidic chip, enabling an unprecedented throughput of up to ∼140 cells/s. To show its broad utility, we use the SRS imaging flow cytometry with the aid of deep learning to study the metabolic heterogeneity of microalgal cells and perform marker-free cancer detection in blood.

Label-Free Chemical Imaging of Latent Fingerprints with Stimulated Raman Scattering Microscopy

2017 ◽  
Vol 89 (8) ◽  
pp. 4468-4473 ◽  
Author(s):  
Benjamin Figueroa ◽  
Yikai Chen ◽  
Kyla Berry ◽  
Andrew Francis ◽  
Dan Fu
Keyword(s):  
Raman Scattering ◽  
Stimulated Raman Scattering ◽  
Chemical Imaging ◽  
Label Free ◽  
Latent Fingerprints ◽  
Stimulated Raman

scholarly journals Stimulated Raman Scattering Microscopy for Label-Free Chemical Imaging

2009 ◽  
Vol 20 (12) ◽  
pp. 30 ◽  
Author(s):  
Christian W. Freudiger ◽  
Wei Min ◽  
Brian G. Saar ◽  
X. Sunney Xie
Keyword(s):  
Raman Scattering ◽  
Stimulated Raman Scattering ◽  
Chemical Imaging ◽  
Label Free ◽  
Stimulated Raman

scholarly journals Super-resolution vibrational imaging using expansion stimulated Raman scattering microscopy

2021 ◽  
Author(s):  
Lixue Shi ◽  
Aleksandra Klimas ◽  
Brendan Gallagher ◽  
Zhangyu Cheng ◽  
Feifei Fu ◽  
...  
Keyword(s):  
Raman Scattering ◽  
Mouse Brain ◽  
Stimulated Raman Scattering ◽  
Super Resolution ◽  
Chemical Imaging ◽  
Metabolic Imaging ◽  
Label Free ◽  
Nanoscale Imaging ◽  
Nanoscale Features ◽  
Stimulated Raman

Stimulated Raman scattering (SRS) microscopy is an emerging technology that provides high chemical specificity for endogenous biomolecules and can circumvent common constraints of fluorescence microscopy including limited capabilities to probe small biomolecules and difficulty resolving many colors simultaneously due to spectral overlap. However, the resolution of SRS microscopy remains governed by the diffraction limit. To overcome this, we describe a new technique called Molecule Anchorable Gel-enabled Nanoscale Imaging of Fluorescence and stImulatEd Raman Scattering microscopy (MAGNIFIERS), that integrates SRS microscopy with expansion microscopy (ExM). ExM is a powerful strategy providing significant improvement in imaging resolution by physical magnification of hydrogel-embedded preserved biological specimens. MAGNIFIERS offers chemical-specific nanoscale imaging with sub-50 nm resolution and has scalable multiplexity when combined with multiplex Raman probes and fluorescent labels. We used MAGNIFIERS to visualize nanoscale features in a label-free manner with C-H vibration of proteins, lipids and DNA in a broad range of biological specimens, from mouse brain, liver and kidney to human lung organoid. In addition, we applied MAGNIFIERS to track nanoscale features of protein synthesis in protein aggregates using metabolic labeling of small metabolites. Finally, we used MAGNIFIERS to demonstrate 8-color nanoscale imaging in an expanded mouse brain section. Overall, MAGNIFIERS is a valuable platform for super-resolution label-free chemical imaging, high-resolution metabolic imaging, and highly multiplexed nanoscale imaging, thus bringing SRS to nanoscopy.

scholarly journals Volumetric chemical imaging by clearing-enhanced stimulated Raman scattering microscopy

2019 ◽  
Vol 116 (14) ◽  
pp. 6608-6617 ◽  
Author(s):  
Mian Wei ◽  
Lingyan Shi ◽  
Yihui Shen ◽  
Zhilun Zhao ◽  
Asja Guzman ◽  
...  
Keyword(s):  
Raman Scattering ◽  
Optical Microscopy ◽  
Stimulated Raman Scattering ◽  
Three Dimensional ◽  
Lipid Synthesis ◽  
Chemical Imaging ◽  
Label Free ◽  
Tissue Clearing ◽  
Metabolic Activities ◽  
Stimulated Raman

Three-dimensional visualization of tissue structures using optical microscopy facilitates the understanding of biological functions. However, optical microscopy is limited in tissue penetration due to severe light scattering. Recently, a series of tissue-clearing techniques have emerged to allow significant depth-extension for fluorescence imaging. Inspired by these advances, we develop a volumetric chemical imaging technique that couples Raman-tailored tissue-clearing with stimulated Raman scattering (SRS) microscopy. Compared with the standard SRS, the clearing-enhanced SRS achieves greater than 10-times depth increase. Based on the extracted spatial distribution of proteins and lipids, our method reveals intricate 3D organizations of tumor spheroids, mouse brain tissues, and tumor xenografts. We further develop volumetric phasor analysis of multispectral SRS images for chemically specific clustering and segmentation in 3D. Moreover, going beyond the conventional label-free paradigm, we demonstrate metabolic volumetric chemical imaging, which allows us to simultaneously map out metabolic activities of protein and lipid synthesis in glioblastoma. Together, these results support volumetric chemical imaging as a valuable tool for elucidating comprehensive 3D structures, compositions, and functions in diverse biological contexts, complementing the prevailing volumetric fluorescence microscopy.

scholarly journals Plasmon-enhanced stimulated Raman scattering microscopy with single-molecule detection sensitivity

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Cheng Zong ◽  
Ranjith Premasiri ◽  
Haonan Lin ◽  
Yimin Huang ◽  
Chi Zhang ◽  
...  
Keyword(s):  
Raman Scattering ◽  
Single Molecule ◽  
High Speed ◽  
Stimulated Raman Scattering ◽  
Chemical Imaging ◽  
Detection Sensitivity ◽  
Single Molecule Detection ◽  
Label Free ◽  
Biomedical Systems ◽  
Stimulated Raman

AbstractStimulated Raman scattering (SRS) microscopy allows for high-speed label-free chemical imaging of biomedical systems. The imaging sensitivity of SRS microscopy is limited to ~10 mM for endogenous biomolecules. Electronic pre-resonant SRS allows detection of sub-micromolar chromophores. However, label-free SRS detection of single biomolecules having extremely small Raman cross-sections (~10−30 cm2 sr−1) remains unreachable. Here, we demonstrate plasmon-enhanced stimulated Raman scattering (PESRS) microscopy with single-molecule detection sensitivity. Incorporating pico-Joule laser excitation, background subtraction, and a denoising algorithm, we obtain robust single-pixel SRS spectra exhibiting single-molecule events, verified by using two isotopologues of adenine and further confirmed by digital blinking and bleaching in the temporal domain. To demonstrate the capability of PESRS for biological applications, we utilize PESRS to map adenine released from bacteria due to starvation stress. PESRS microscopy holds the promise for ultrasensitive detection and rapid mapping of molecular events in chemical and biomedical systems.

scholarly journals Utilizing Stimulated Raman Scattering Microscopy To Study Intracellular Distribution of Label-Free Ponatinib in Live Cells

2019 ◽  
Vol 63 (5) ◽  
pp. 2028-2034 ◽  
Author(s):  
Kristel Sepp ◽  
Martin Lee ◽  
Marie T. J. Bluntzer ◽  
G. Vignir Helgason ◽  
Alison N. Hulme ◽  
...  
Keyword(s):  
Raman Scattering ◽  
Stimulated Raman Scattering ◽  
Intracellular Distribution ◽  
Live Cells ◽  
Label Free ◽  
Stimulated Raman
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