Exploring Morphological and Biochemical Linkages in Fungal Growth with Label-Free Light Sheet Microscopy and Raman Spectroscopy

ChemPhysChem ◽  
2016 ◽  
Vol 18 (1) ◽  
pp. 72-78 ◽  
Author(s):  
Soumik Siddhanta ◽  
Santosh Kumar Paidi ◽  
Kathryn Bushley ◽  
Ram Prasad ◽  
Ishan Barman
Keyword(s):  
Raman Spectroscopy ◽  
Fungal Growth ◽  
Light Sheet ◽  
Label Free ◽  
Light Sheet Microscopy

scholarly journals Back Cover: Exploring Morphological and Biochemical Linkages in Fungal Growth with Label-Free Light Sheet Microscopy and Raman Spectroscopy (ChemPhysChem 1/2017)

ChemPhysChem ◽  
2017 ◽  
Vol 18 (1) ◽  
pp. 172-172
Author(s):  
Soumik Siddhanta ◽  
Santosh Kumar Paidi ◽  
Kathryn Bushley ◽  
Ram Prasad ◽  
Ishan Barman
Keyword(s):  
Raman Spectroscopy ◽  
Fungal Growth ◽  
Light Sheet ◽  
Label Free ◽  
Back Cover ◽  
Light Sheet Microscopy

scholarly journals Label-free and Multimodal Second Harmonic Generation Light Sheet Microscopy

2020 ◽  
Author(s):  
Niall Hanrahan ◽  
Simon I. R. Lane ◽  
Peter Johnson ◽  
Konstantinos Bourdakos ◽  
Christopher Brereton ◽  
...  
Keyword(s):  
Second Harmonic Generation ◽  
Harmonic Generation ◽  
Biological Samples ◽  
3D Imaging ◽  
Second Harmonic ◽  
Three Dimensional ◽  
Light Sheet ◽  
Optical Methods ◽  
Label Free ◽  
Light Sheet Microscopy

AbstractLight sheet microscopy (LSM) has emerged as one of most profound three dimensional (3D) imaging tools in the life sciences over the last decade. However, LSM is currently performed with fluorescence detection on one- or multi-photon excitation. Label-free LSM imaging approaches have been rather limited. Second Harmonic Generation (SHG) imaging is a label-free technique that has enabled detailed investigation of collagenous structures, including its distribution and remodelling in cancers and respiratory tissue, and how these link to disease. SHG is generally regarded as having only forward- and back-scattering components, apparently precluding the orthogonal detection geometry used in Light Sheet Microscopy. In this work we demonstrate SHG imaging on a light sheet microscope (SHG-LSM) using a rotated Airy beam configuration that demonstrates a powerful new approach to direct, without any further processing or deconvolution, 3D imaging of harmonophores such as collagen in biological samples. We provide unambiguous identification of SHG signals on the LSM through its wavelength and polarisation sensitivity. In a multimodal LSM setup we demonstrate that SHG and two-photon signals can be acquired on multiple types of different biological samples. We further show that SHG-LSM is sensitive to changes in collagen synthesis within lung fibroblast 3D cell cultures. This work expands on the existing optical methods available for use with light sheet microscopy, adding a further label-free imaging technique which can be combined with other detection modalities to realise a powerful multi-modal microscope for 3D bioimaging.

scholarly journals Integrative quantitative-phase and airy light-sheet imaging

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
N. R. Subedi ◽  
P. S. Jung ◽  
E. L. Bredeweg ◽  
S. Nemati ◽  
S. E. Baker ◽  
...  
Keyword(s):  
Evolutionary Biology ◽  
Imaging System ◽  
Light Sheet ◽  
Live Cells ◽  
Phase Imaging ◽  
Label Free ◽  
Quantitative Phase Imaging ◽  
Quantitative Phase ◽  
Light Sheet Microscopy ◽  
Order Of Magnitude

AbstractLight-sheet microscopy enables considerable speed and phototoxicity gains, while quantitative-phase imaging confers label-free recognition of cells and organelles, and quantifies their number-density that, thermodynamically, is more representative of metabolism than size. Here, we report the fusion of these two imaging modalities onto a standard inverted microscope that retains compatibility with microfluidics and open-source software for image acquisition and processing. An accelerating Airy-beam light-sheet critically enabled imaging areas that were greater by more than one order of magnitude than a Gaussian beam illumination and matched exactly those of quantitative-phase imaging. Using this integrative imaging system, we performed a demonstrative multivariate investigation of live-cells in microfluidics that unmasked that cellular noise can affect the compartmental localization of metabolic reactions. We detail the design, assembly, and performance of the integrative imaging system, and discuss potential applications in biotechnology and evolutionary biology.

scholarly journals Multiview deconvolution approximation multiphoton microscopy of tissues and zebrafish larvae

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Dimitrios Kapsokalyvas ◽  
Rodrigo Rosas ◽  
Rob W. A. Janssen ◽  
Jo M. Vanoevelen ◽  
Miranda Nabben ◽  
...  
Keyword(s):  
Second Harmonic ◽  
Multiphoton Microscopy ◽  
Light Sheet ◽  
Three Dimensions ◽  
Fluorescence Signal ◽  
Label Free ◽  
Microscopy Imaging ◽  
Isotropic Resolution ◽  
Light Sheet Microscopy ◽  
3D Image Reconstruction

AbstractImaging in three dimensions is necessary for thick tissues and small organisms. This is possible with tomographic optical microscopy techniques such as confocal, multiphoton and light sheet microscopy. All these techniques suffer from anisotropic resolution and limited penetration depth. In the past, Multiview microscopy—imaging the sample from different angles followed by 3D image reconstruction—was developed to address this issue for light sheet microscopy based on fluorescence signal. In this study we applied this methodology to accomplish Multiview imaging with multiphoton microscopy based on fluorescence and additionally second harmonic signal from myosin and collagen. It was shown that isotropic resolution was achieved, the entirety of the sample was visualized, and interference artifacts were suppressed allowing clear visualization of collagen fibrils and myofibrils. This method can be applied to any scanning microscopy technique without microscope modifications. It can be used for imaging tissue and whole mount small organisms such as heart tissue, and zebrafish larva in 3D, label-free or stained, with at least threefold axial resolution improvement which can be significant for the accurate quantification of small 3D structures.

Second Harmonic Generation Light Sheet Microscopy (SHG-LSM): A new tool for label-free 3D bioimaging

2020 ◽  
Author(s):  
Niall Hanrahan ◽  
Simon Lane ◽  
Peter Johnson ◽  
Konstantinos Bourdakos ◽  
Christopher J. Brereton ◽  
...  
Keyword(s):  
Second Harmonic Generation ◽  
Harmonic Generation ◽  
Second Harmonic ◽  
Light Sheet ◽  
Label Free ◽  
Light Sheet Microscopy

scholarly journals Visualization and Analysis of Whole Depot Adipose Tissue Innervation

2019 ◽  
Author(s):  
Jake W. Willows ◽  
Magdalena Blaszkiewicz ◽  
Amy Lamore ◽  
Samuel Borer ◽  
Amanda L. Dubois ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Second Harmonic ◽  
Imaging Techniques ◽  
Three Dimensional ◽  
Light Sheet ◽  
Tissue Imaging ◽  
Label Free ◽  
Adipose Tissues ◽  
Light Sheet Microscopy ◽  
Neurite Density

AbstractAdipose tissue requires neural innervation in order to regulate important metabolic functions. Though seminal work on adipose denervation has underscored the importance of adipose-nerve interactions in both white (energy storing) and brown (energy expending) adipose tissues, much remains a mystery. This is due, in part, to the inability to effectively visualize the various nerve subtypes residing within these tissues and to gain a comprehensive quantitation of neurite density in an entire depot. With the recent surge of advanced imaging techniques such as light sheet microscopy and optical clearing procedures, adipose tissue imaging has been reinvigorated with a focus on three-dimensional analysis of tissue innervation. However, clearing techniques are time consuming, often require solvents caustic to objective lenses, alter tissue morphology, and greatly reduce fluorophore lifespan. Not only are current methods of imaging wholemount adipose tissues inconvenient, but often attempts to quantify neurite density across physiological or pathophysiological conditions have been limited to representative section sampling. We have developed a new method of adipose tissue neurite imaging and quantitation that is faster than current clearing-based methods, does not require caustic chemicals, and leaves the tissue fully intact. Maintenance of a fully intact depot allowed for tiling z-stacks and producing maximum intensity projections of the entire adipose depot, which were then used to quantify neurite density across the tissue. With this processing method we were able to characterize the nerves, nerve-subtypes, and neurovascular interactions within the inguinal subcutaneous white adipose tissue in mice using up to five fluorescent channels at high resolution. We also utilized second harmonic generation, which provides label-free imaging, to investigate collagen fiber abundance in adipose of obese mice.

scholarly journals Autofluorescence of stingray skeletal cartilage: hyperspectral imaging as a tool for histological characterization

2021 ◽  
Vol 1 (1) ◽  
Author(s):  
Júlia Chaumel ◽  
María Marsal ◽  
Adrián Gómez-Sánchez ◽  
Michael Blumer ◽  
Emilio J. Gualda ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Light Sheet ◽  
Label Free ◽  
Cell Nuclei ◽  
Micro Computed Tomography ◽  
Composite Tissue ◽  
Light Sheet Microscopy ◽  
Imaging Tool ◽  
Biochemical Signals ◽  
First Time

AbstractTessellated cartilage is a distinctive composite tissue forming the bulk of the skeleton of cartilaginous fishes (e.g. sharks and rays), built from unmineralized cartilage covered at the surface by a thin layer of mineralized tiles called tesserae. The finescale structure and composition of elasmobranch tessellated cartilage has largely been investigated with electron microscopy, micro-computed tomography and histology, but many aspects of tissue structure and composition remain uncharacterized. In our study, we demonstrate that the tessellated cartilage of a stingray exhibits a strong and diverse autofluorescence, a native property of the tissue which can be harnessed as an effective label-free imaging technique. The autofluorescence signal was excited using a broad range of wavelengths in confocal and light sheet microscopy, comparing several sample preparations (fresh; demineralized and paraffin-embedded; non-demineralized and plastic-embedded) and imaging the tissue at different scales. Autofluorescence varied with sample preparation with the signal in both plastic- and paraffin-embedded samples strong enough to allow visualization of finescale (≥ 1 μm) cellular and matrix structures, such as cell nuclei and current and former mineralization fronts, identifiable by globular mineralized tissue. A defined pericellular matrix (PCM) surrounding chondrocytes was also discernible, described here for the first time in elasmobranchs. The presence of a PCM suggests similarities with mammalian cartilage regarding how chondrocytes interact with their environment, the PCM in mammals acting as a transducer for biomechanical and biochemical signals. A posterior analysis of hyperspectral images by an MCR-ALS unmixing algorithm allowed identification of several distinct fluorescence signatures associated to specific regions in the tissue. Some fluorescence signatures identified could be correlated with collagen type II, the most abundant structural molecule of cartilage. Other fluorescence signatures, however, remained unidentified, spotlighting tissue regions that deserve deeper characterization and suggesting the presence of molecules still unidentified in elasmobranch skeletal cartilage. Our results show that autofluorescence can be a powerful exploratory imaging tool for characterizing less-studied skeletal tissues, such as tessellated cartilage. The images obtained are largely comparable with more commonly used techniques, but without the need for complicated sample preparations or external staining reagents standard in histology and electron microscopy (TEM, SEM).

scholarly journals Achromatic terahertz Airy beam generation with dielectric metasurfaces

Nanophotonics ◽  
2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Qingqing Cheng ◽  
Juncheng Wang ◽  
Ling Ma ◽  
Zhixiong Shen ◽  
Jing Zhang ◽  
...  
Keyword(s):  
Biomedical Imaging ◽  
Frequency Dispersion ◽  
Light Sheet ◽  
Photonic Devices ◽  
Self Healing ◽  
Beam Focusing ◽  
Light Sheet Microscopy ◽  
Airy Beam ◽  
Potential Applications ◽  
Airy Beams

AbstractAiry beams exhibit intriguing properties such as nonspreading, self-bending, and self-healing and have attracted considerable recent interest because of their many potential applications in photonics, such as to beam focusing, light-sheet microscopy, and biomedical imaging. However, previous approaches to generate Airy beams using photonic structures have suffered from severe chromatic problems arising from strong frequency dispersion of the scatterers. Here, we design and fabricate a metasurface composed of silicon posts for the frequency range 0.4–0.8 THz in transmission mode, and we experimentally demonstrate achromatic Airy beams exhibiting autofocusing properties. We further show numerically that a generated achromatic Airy-beam-based metalens exhibits self-healing properties that are immune to scattering by particles and that it also possesses a larger depth of focus than a traditional metalens. Our results pave the way to the realization of flat photonic devices for applications to noninvasive biomedical imaging and light-sheet microscopy, and we provide a numerical demonstration of a device protocol.

scholarly journals Single-Molecule Light-Sheet Microscopy with Local Nanopipette Delivery

2021 ◽  
Vol 93 (8) ◽  
pp. 4092-4099
Author(s):  
Bing Li ◽  
Aleks Ponjavic ◽  
Wei-Hsin Chen ◽  
Lee Hopkins ◽  
Craig Hughes ◽  
...  
Keyword(s):  
Single Molecule ◽  
Light Sheet ◽  
Light Sheet Microscopy
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