scholarly journals Depth-resolved mid-infrared photothermal imaging of living cells and organisms with submicrometer spatial resolution

2016 ◽  
Vol 2 (9) ◽  
pp. e1600521 ◽  
Author(s):  
Delong Zhang ◽  
Chen Li ◽  
Chi Zhang ◽  
Mikhail N. Slipchenko ◽  
Gregory Eakins ◽  
...  
Keyword(s):  
Spatial Resolution ◽  
Single Cells ◽  
Chemical Imaging ◽  
Detection Sensitivity ◽  
Living Systems ◽  
Live Cells ◽  
Photothermal Effect ◽  
Label Free ◽  
Infrared Microscopy ◽  
Mid Infrared

Chemical contrast has long been sought for label-free visualization of biomolecules and materials in complex living systems. Although infrared spectroscopic imaging has come a long way in this direction, it is thus far only applicable to dried tissues because of the strong infrared absorption by water. It also suffers from low spatial resolution due to long wavelengths and lacks optical sectioning capabilities. We overcome these limitations through sensing vibrational absorption–induced photothermal effect by a visible laser beam. Our mid-infrared photothermal (MIP) approach reached 10 μM detection sensitivity and submicrometer lateral spatial resolution. This performance has exceeded the diffraction limit of infrared microscopy and allowed label-free three-dimensional chemical imaging of live cells and organisms. Distributions of endogenous lipid and exogenous drug inside single cells were visualized. We further demonstrated in vivo MIP imaging of lipids and proteins inCaenorhabditis elegans. The reported MIP imaging technology promises broad applications from monitoring metabolic activities to high-resolution mapping of drug molecules in living systems, which are beyond the reach of current infrared microscopy.

Label-free biochemical refractive-index tomography of single cells with mid-infrared photothermal effect

2020 ◽  
Author(s):  
Miu Tamamitsu ◽  
Keiichiro Toda ◽  
Hiroyuki Shimada ◽  
Yu Nagashima ◽  
Ryoichi Horisaki ◽  
...  
Keyword(s):  
Refractive Index ◽  
Single Cells ◽  
Photothermal Effect ◽  
Label Free ◽  
Mid Infrared

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.

Tracking the Formation and Degradation of Fatty-Acid-Accumulated Mitochondria Using Label-Free Chemical Imaging

2020 ◽  
Author(s):  
Chi Zhang ◽  
Stephen Boppart
Keyword(s):  
Fatty Acid ◽  
Structural Features ◽  
Chemical Imaging ◽  
Live Cells ◽  
Label Free ◽  
Dynamic Changes ◽  
Regulatory Processes ◽  
Transport Chain ◽  
Anti Stokes ◽  
First Time

Abstract The mitochondrion is one of the key organelles for maintaining cellular homeostasis. External environmental stimuli and internal regulatory processes alter the metabolism and functions of mitochondria. To understand these activities of mitochondria, it is critical to probe the key metabolic molecules inside these organelles. In this study, we used label-free chemical imaging modalities including coherent anti-Stokes Raman scattering and multiphoton-excited autofluorescence to study the mitochondrial activities in living cancer cells. We found that hypothermia exposure tends to induce fatty-acid (FA) accumulation in some mitochondria of MIAPaCa-2 cells. Autofluorescence images show that the FA-accumulated mitochondria also have abnormal NADH and FAD metabolism, likely induced by the dysfunction of the electron transport chain. We also found that when the cells were re-warmed to physiological temperature after a period of hypothermia, the FA-accumulated mitochondria changed their structural features, likely caused by the mitophagy process. To the best of our knowledge, this is the first time that FA accumulation in mitochondria was observed in live cells. Our research also demonstrates that multimodal label-free chemical imaging is an attractive tool to discover abnormal functions of mitochondria at the single-organelle level and can be used to quantify the dynamic changes of this organelle under perturbative conditions.

scholarly journals Fibronectin-Based Nanomechanical Biosensors to Map 3D Strains in Live Cells and Tissues

2020 ◽  
Author(s):  
Daniel J. Shiwarski ◽  
Joshua W. Tashman ◽  
Alkiviadis Tsamis ◽  
Jacqueline M. Bliley ◽  
Malachi A. Blundon ◽  
...  
Keyword(s):  
Spatial Resolution ◽  
Single Cells ◽  
Time Lapse ◽  
Confocal Imaging ◽  
Square Lattice ◽  
Live Cells ◽  
3D Analysis ◽  
Wide Range ◽  
And Function ◽  
Analysis Platform

AbstractMechanical forces are integral to a wide range of cellular processes including migration, differentiation and tissue morphogenesis; however, it has proved challenging to directly measure strain at high spatial resolution and with minimal tissue perturbation. Here, we fabricated, calibrated, and tested a fibronectin (FN)-based nanomechanical biosensor (NMBS) that can be applied to cells and tissues to measure the magnitude, direction, and dynamics of strain from subcellular to tissue length-scales. The NMBS is a fluorescently-labeled, ultrathin square lattice FN mesh with spatial resolution tailored by adjusting the width and spacing of the lattice fibers from 2-100 µm. Time-lapse 3D confocal imaging of the NMBS demonstrated strain tracking in 2D and 3D following mechanical deformation of known materials and was validated with finite element modeling. Imaging and 3D analysis of the NMBS applied to single cells, cell monolayers, and Drosophila ovarioles demonstrated the ability to dynamically track microscopic tensile and compressive strains in various biological applications with minimal tissue perturbation. This fabrication and analysis platform serves as a novel tool for studying cells, tissues, and more complex systems where forces guide structure and function.

scholarly journals Mid-infrared optoacoustic microscopy with label-free chemical contrast in living cells and tissues

2018 ◽  
Author(s):  
Miguel A. Pleitez ◽  
Asrar Ali Khan ◽  
Josefine Reber ◽  
Andriy Chmyrov ◽  
Markus R. Seeger ◽  
...  
Keyword(s):  
Imaging Modality ◽  
Living Cells ◽  
Spatiotemporal Dynamics ◽  
Live Cell ◽  
Imaging Modalities ◽  
Label Free ◽  
Infrared Microscopy ◽  
Mid Infrared ◽  
Infrared Sensing ◽  
Biomolecular Imaging

We developed mid-infrared optoacoustic microscopy (MiROM), a bond-selective imaging modality that overcomes water/tissue opacity and depth limitations of mid-infrared sensing allowing uncompromised live-cell/thick-tissue mid-infrared microscopy with up to three orders of magnitudehigher sensitivity than other vibrational imaging modalities; such as Raman. We showcase the functional label-free biomolecular imaging capabilities of MiROM by monitoring the spatiotemporal dynamics of lipids and proteins during lipolysis in living adipocytes. Since MiROM, contrary to Ramanmodalities, is not only able to detect lipids and proteins, but also important metabolites such as glucose without the need of labels, here we discuss how MiROM yields novel functional label-free abilities for a broader range of analytical studies in living cells and tissues.

scholarly journals Inside Cover: Label-Free Chemical Imaging of Catalytic Solids by Coherent Anti-Stokes Raman Scattering and Synchrotron-Based Infrared Microscopy (Angew. Chem. Int. Ed. 47/2009)

2009 ◽  
Vol 48 (47) ◽  
pp. 8798-8798
Author(s):  
Marianne���H.���F. Kox ◽  
Katrin���F. Domke ◽  
James���P.���R. Day ◽  
Gianluca Rago ◽  
Eli Stavitski ◽  
...  
Keyword(s):  
Raman Scattering ◽  
Chemical Imaging ◽  
Label Free ◽  
Infrared Microscopy ◽  
Anti Stokes

scholarly journals High-throughput label-free molecular fingerprinting flow cytometry

2019 ◽  
Vol 5 (1) ◽  
pp. eaau0241 ◽  
Author(s):  
Kotaro Hiramatsu ◽  
Takuro Ideguchi ◽  
Yusuke Yonamine ◽  
SangWook Lee ◽  
Yizhi Luo ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Single Cell ◽  
High Throughput ◽  
Single Cell Analysis ◽  
Single Cells ◽  
Fluorescent Labeling ◽  
Live Cells ◽  
Molecular Fingerprinting ◽  
Label Free ◽  
Indispensable Tool

Flow cytometry is an indispensable tool in biology for counting and analyzing single cells in large heterogeneous populations. However, it predominantly relies on fluorescent labeling to differentiate cells and, hence, comes with several fundamental drawbacks. Here, we present a high-throughput Raman flow cytometer on a microfluidic chip that chemically probes single live cells in a label-free manner. It is based on a rapid-scan Fourier-transform coherent anti-Stokes Raman scattering spectrometer as an optical interrogator, enabling us to obtain the broadband molecular vibrational spectrum of every single cell in the fingerprint region (400 to 1600 cm−1) with a record-high throughput of ~2000 events/s. As a practical application of the method not feasible with conventional flow cytometry, we demonstrate high-throughput label-free single-cell analysis of the astaxanthin productivity and photosynthetic dynamics ofHaematococcus lacustris.

Label-Free Chemical Imaging of Catalytic Solids by Coherent Anti-Stokes Raman Scattering and Synchrotron-Based Infrared Microscopy

2009 ◽  
Vol 121 (47) ◽  
pp. 9152-9156 ◽  
Author(s):  
Marianne���H.���F. Kox ◽  
Katrin���F. Domke ◽  
James���P.���R. Day ◽  
Gianluca Rago ◽  
Eli Stavitski ◽  
...  
Keyword(s):  
Raman Scattering ◽  
Chemical Imaging ◽  
Label Free ◽  
Infrared Microscopy ◽  
Anti Stokes

Advancements in quantum cascade laser-based infrared microscopy of aqueous media

2016 ◽  
Vol 187 ◽  
pp. 119-134 ◽  
Author(s):  
K. Haase ◽  
N. Kröger-Lui ◽  
A. Pucci ◽  
A. Schönhals ◽  
W. Petrich
Keyword(s):  
Quantum Cascade Laser ◽  
Quantum Cascade Lasers ◽  
Spectral Power ◽  
Aqueous Media ◽  
Aqueous Environment ◽  
Label Free ◽  
Infrared Microscopy ◽  
Signal To Noise ◽  
Mid Infrared ◽  
Quantum Cascade

The large mid-infrared absorption coefficient of water frequently hampers the rapid, label-free infrared microscopy of biological objects in their natural aqueous environment. However, the high spectral power density of quantum cascade lasers is shifting this limitation such that mid-infrared absorbance images can be acquired in situ within signal-to-noise ratios of up to 100. Even at sample thicknesses well above 50 μm, signal-to-noise ratios above 10 are readily achieved. The quantum cascade laser-based microspectroscopy of aqueous media is exemplified by imaging an aqueous yeast solution and quantifying glucose consumption, ethanol generation as well as the production of carbon dioxide gas during fermentation.

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