Label-free chemical imaging of live Euglena gracilis by high-speed SRS spectral microscopy (Conference Presentation)

2016 ◽  
Author(s):  
Yoshifumi Wakisaka ◽  
Yuta Suzuki ◽  
Kyoya Tokunaga ◽  
Misa Hirose ◽  
Ryota Domon ◽  
...  
Keyword(s):  
Euglena Gracilis ◽  
High Speed ◽  
Chemical Imaging ◽  
Label Free

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.

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 High-speed label-free two-photon fluorescence microscopy of metabolic transients during neuronal activity

2021 ◽  
Vol 118 (8) ◽  
pp. 081104
Author(s):  
Andrew J. Bower ◽  
Carlos Renteria ◽  
Joanne Li ◽  
Marina Marjanovic ◽  
Ronit Barkalifa ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Neuronal Activity ◽  
High Speed ◽  
Label Free ◽  
Two Photon ◽  
Metabolic Transients ◽  
Two Photon Fluorescence ◽  
Photon Fluorescence

scholarly journals Label Free High Speed Wide Field Imaging of Single Microtubules using Interference Reflection Microscopy

2018 ◽  
Vol 114 (3) ◽  
pp. 504a-505a
Author(s):  
Mohammed Mahamdeh ◽  
Steve Simmert ◽  
Anna Łuchniak ◽  
Erik Schäffer ◽  
Jonathon Howard
Keyword(s):  
High Speed ◽  
Wide Field ◽  
Label Free ◽  
Field Imaging ◽  
Wide Field Imaging

scholarly journals Rapid Antibody Selection Using Surface Plasmon Resonance for High-Speed and Sensitive Hazelnut Lateral Flow Prototypes

Biosensors ◽  
2018 ◽  
Vol 8 (4) ◽  
pp. 130 ◽  
Author(s):  
Georgina Ross ◽  
Maria Bremer ◽  
Jan Wichers ◽  
Aart van Amerongen ◽  
Michel Nielen
Keyword(s):  
Surface Plasmon Resonance ◽  
Surface Plasmon ◽  
Plasmon Resonance ◽  
High Speed ◽  
Selection Process ◽  
Low Cost ◽  
Real Life ◽  
Cross Reactivity ◽  
Lateral Flow ◽  
Label Free

Lateral Flow Immunoassays (LFIAs) allow for rapid, low-cost, screening of many biomolecules such as food allergens. Despite being classified as rapid tests, many LFIAs take 10–20 min to complete. For a really high-speed LFIA, it is necessary to assess antibody association kinetics. By using a label-free optical technique such as Surface Plasmon Resonance (SPR), it is possible to screen crude monoclonal antibody (mAb) preparations for their association rates against a target. Herein, we describe an SPR-based method for screening and selecting crude anti-hazelnut antibodies based on their relative association rates, cross reactivity and sandwich pairing capabilities, for subsequent application in a rapid ligand binding assay. Thanks to the SPR selection process, only the fast mAb (F-50-6B12) and the slow (S-50-5H9) mAb needed purification for labelling with carbon nanoparticles to exploit high-speed LFIA prototypes. The kinetics observed in SPR were reflected in LFIA, with the test line appearing within 30 s, almost two times faster when F-50-6B12 was used, compared with S-50-5H9. Additionally, the LFIAs have demonstrated their future applicability to real life samples by detecting hazelnut in the sub-ppm range in a cookie matrix. Finally, these LFIAs not only provide a qualitative result when read visually, but also generate semi-quantitative data when exploiting freely downloadable smartphone apps.

scholarly journals Viewing Cancer in a New Light: Multimodal Label-Free Specific Chemical Imaging to Distinguish Healthy Cell Tissue from Invasive Carcinoma

2017 ◽  
Vol 112 (3) ◽  
pp. 187a
Author(s):  
Bjarne Thorsted ◽  
Stine R. Larsen ◽  
Christian Godballe ◽  
Jonathan R. Brewer
Keyword(s):  
Invasive Carcinoma ◽  
Chemical Imaging ◽  
Label Free ◽  
Specific Chemical ◽  
Healthy Cell ◽  
Cell Tissue

scholarly journals Whole organ cross-section chemical imaging using label-free mega-mosaic FTIR microscopy

The Analyst ◽  
2013 ◽  
Vol 138 (23) ◽  
pp. 7066 ◽  
Author(s):  
Paul Bassan ◽  
Ashwin Sachdeva ◽  
Jonathan H. Shanks ◽  
Mick D. Brown ◽  
Noel W. Clarke ◽  
...  
Keyword(s):  
Cross Section ◽  
Chemical Imaging ◽  
Label Free ◽  
Ftir Microscopy

Monitoring the effects of chemical stimuli on live cells with metasurface-enhanced infrared reflection spectroscopy

2021 ◽  
Author(s):  
Steven H. Huang ◽  
Jiaruo Li ◽  
Zhiyuan Fan ◽  
Robert Delgado ◽  
Gennady Shvets
Keyword(s):  
Intracellular Signaling ◽  
Strong Field ◽  
Detection System ◽  
Field Enhancement ◽  
Chemical Imaging ◽  
Live Cells ◽  
Label Free ◽  
Reflection Spectroscopy ◽  
Infrared Reflection ◽  
Infrared Reflection Spectroscopy

Infrared spectroscopy has found wide applications in the analysis of biological materials. A more recent development is the use of engineered nanostructures, or plasmonic metasurfaces, as substrates for metasurface-enhanced infrared reflection spectroscopy (MEIRS). Here, we demonstrate that strong field enhancement from plasmonic metasurfaces enables the use of MEIRS as a highly informative analytic technique for real-time monitoring of cells. By exposing live cells cultured on a plasmonic metasurface to chemical compounds, we show that MEIRS can be used as a label-free phenotypic assay for detecting multiple cellular responses to external stimuli: changes in cell morphology, adhesion, lipid composition of the cellular membrane, as well as intracellular signaling. Using a focal plane array detection system, we show that MEIRS also enables spectro-chemical imaging at the single-cell level. The described metasurface-based all-optical sensor opens the way to a scalable, high-throughput spectroscopic assay for live cells.

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