scholarly journals Label-free super-resolution chemical imaging of biomedical specimens

2021 ◽  
Author(s):  
Julien Guilbert ◽  
Awoke Negash ◽  
Simon Labouesse ◽  
Sylvain Gigan ◽  
Anne Sentenac ◽  
...  
Keyword(s):  
Stimulated Raman Scattering ◽  
Super Resolution ◽  
Chemical Imaging ◽  
Tissue Imaging ◽  
Label Free ◽  
Structured Illumination ◽  
Resolution Method ◽  
Fast Acquisition ◽  
Chemically Selective ◽  
Single Pixel

Raman microscopy provides chemically selective imaging by exploiting intrinsic vibrational properties of specimens. Yet, a fast acquisition, low phototoxicity, and non-specific (to a vibrational/electronic mode) super-resolution method has been elusive for tissue imaging. We demonstrate a single-pixel-based approach, combined with robust structured illumination, that enables fast super-resolution in stimulated Raman scattering microscopy at low power levels. The methodology is straightforward to implement and compatible with thick biological specimens, therefore paving the way for probing complex biological systems when exogenous labelling is challenging.

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 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 fluorescence microscopy by line-scanning with an unmodified two-photon microscope

2021 ◽  
Vol 379 (2199) ◽  
Author(s):  
Christian Pilger ◽  
Jakub Pospíšil ◽  
Marcel Müller ◽  
Martin Ruoff ◽  
Martin Schütte ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Super Resolution ◽  
Image Resolution ◽  
Laser Scanning Microscopy ◽  
Tissue Imaging ◽  
Source Image ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Fluorescence Excitation ◽  
Two Photon

Fluorescence-based microscopy as one of the standard tools in biomedical research benefits more and more from super-resolution methods, which offer enhanced spatial resolution allowing insights into new biological processes. A typical drawback of using these methods is the need for new, complex optical set-ups. This becomes even more significant when using two-photon fluorescence excitation, which offers deep tissue imaging and excellent z-sectioning. We show that the generation of striped-illumination patterns in two-photon laser scanning microscopy can readily be exploited for achieving optical super-resolution and contrast enhancement using open-source image reconstruction software. The special appeal of this approach is that even in the case of a commercial two-photon laser scanning microscope no optomechanical modifications are required to achieve this modality. Modifying the scanning software with a custom-written macro to address the scanning mirrors in combination with rapid intensity switching by an electro-optic modulator is sufficient to accomplish the acquisition of two-photon striped-illumination patterns on an sCMOS camera. We demonstrate and analyse the resulting resolution improvement by applying different recently published image resolution evaluation procedures to the reconstructed filtered widefield and super-resolved images. This article is part of the Theo Murphy meeting issue ‘Super-resolution structured illumination microscopy (part 1)'.

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 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.

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