scholarly journals A Multimodal Adaptive Super-Resolution and Confocal Microscope

2018 ◽  
Author(s):  
Liyana Valiya Peedikakkal ◽  
Andrew Furley ◽  
Ashley J. Cadby
Keyword(s):  
Imaging System ◽  
Imaging Modality ◽  
Super Resolution ◽  
Single Mode ◽  
Imaging Modalities ◽  
Structured Illumination ◽  
Wide Range ◽  
Transition Times ◽  
Localisation Microscopy ◽  
Microscopy Techniques

Existing optical microscopy techniques compromise between resolution, photodamage, speed of acquisition and imaging in to deep samples. This often confines a technique to a certain biological system or process. We present a versatile imaging system which can switch between imaging modalities with sub millisecond transition times to adapt to the needs of a wide range of sample types. The imaging modalities provide the minimally invasive but low-resolution epi-fluorescence though increasing invasive but higher resolution confocal and structured illumination until the highest resolution is achieved through the most intrusive, localisation microscopy. The ability of the system to overcome the limitations of conventional single mode microscopy is demonstrated by several biological investigations. The ideas presented in this work allow researchers to move away from the model of a single imaging modality to study a specific process and instead follow those processes using the most suitable method available during the lifetime of the investigation.

scholarly journals Extended mechanical force measurements using structured illumination microscopy

2021 ◽  
Vol 379 (2199) ◽  
Author(s):  
Kseniya Korobchevskaya ◽  
Huw Colin-York ◽  
Liliana Barbieri ◽  
Marco Fritzsche
Keyword(s):  
Imaging System ◽  
Three Dimensional ◽  
Traction Force ◽  
Super Resolution ◽  
Mechanical Force ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Biological Studies ◽  
Wide Range ◽  
Spatio Temporal

Quantifying cell generated mechanical forces is key to furthering our understanding of mechanobiology. Traction force microscopy (TFM) is one of the most broadly applied force probing technologies, but its sensitivity is strictly dependent on the spatio-temporal resolution of the underlying imaging system. In previous works, it was demonstrated that increased sampling densities of cell derived forces permitted by super-resolution fluorescence imaging enhanced the sensitivity of the TFM method. However, these recent advances to TFM based on super-resolution techniques were limited to slow acquisition speeds and high illumination powers. Here, we present three novel TFM approaches that, in combination with total internal reflection, structured illumination microscopy and astigmatism, improve the spatial and temporal performance in either two-dimensional or three-dimensional mechanical force quantification, while maintaining low illumination powers. These three techniques can be straightforwardly implemented on a single optical set-up offering a powerful platform to provide new insights into the physiological force generation in a wide range of biological studies. This article is part of the Theo Murphy meeting issue ‘Super-resolution structured illumination microscopy (part 1)'.

scholarly journals Double moiré localized plasmon structured illumination microscopy

Nanophotonics ◽  
2020 ◽  
Vol 0 (0) ◽  
Author(s):  
Ruslan Röhrich ◽  
A. Femius Koenderink
Keyword(s):  
Near Field ◽  
Super Resolution ◽  
Reconstruction Algorithm ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Experimental Realization ◽  
Spatial Frequencies ◽  
Wide Range ◽  
Localized Plasmon ◽  
Plasmonic Grating

AbstractStructured illumination microscopy (SIM) is a well-established fluorescence imaging technique, which can increase spatial resolution by up to a factor of two. This article reports on a new way to extend the capabilities of structured illumination microscopy, by combining ideas from the fields of illumination engineering and nanophotonics. In this technique, plasmonic arrays of hexagonal symmetry are illuminated by two obliquely incident beams originating from a single laser. The resulting interference between the light grating and plasmonic grating creates a wide range of spatial frequencies above the microscope passband, while still preserving the spatial frequencies of regular SIM. To systematically investigate this technique and to contrast it with regular SIM and localized plasmon SIM, we implement a rigorous simulation procedure, which simulates the near-field illumination of the plasmonic grating and uses it in the subsequent forward imaging model. The inverse problem, of obtaining a super-resolution (SR) image from multiple low-resolution images, is solved using a numerical reconstruction algorithm while the obtained resolution is quantitatively assessed. The results point at the possibility of resolution enhancements beyond regular SIM, which rapidly vanishes with the height above the grating. In an initial experimental realization, the existence of the expected spatial frequencies is shown and the performance of compatible reconstruction approaches is compared. Finally, we discuss the obstacles of experimental implementations that would need to be overcome for artifact-free SR imaging.

scholarly journals Super-resolution imaging of fluorescent dipoles via polarized structured illumination microscopy

2019 ◽  
Vol 10 (1) ◽  
Author(s):  
Karl Zhanghao ◽  
Xingye Chen ◽  
Wenhui Liu ◽  
Meiqi Li ◽  
Yiqiong Liu ◽  
...  
Keyword(s):  
Hippocampal Neurons ◽  
Fluorescent Protein ◽  
Imaging Modality ◽  
Super Resolution ◽  
Vital Role ◽  
Molecular Structures ◽  
Polarization Microscopy ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Resolution Imaging

Abstract Fluorescence polarization microscopy images both the intensity and orientation of fluorescent dipoles and plays a vital role in studying molecular structures and dynamics of bio-complexes. However, current techniques remain difficult to resolve the dipole assemblies on subcellular structures and their dynamics in living cells at super-resolution level. Here we report polarized structured illumination microscopy (pSIM), which achieves super-resolution imaging of dipoles by interpreting the dipoles in spatio-angular hyperspace. We demonstrate the application of pSIM on a series of biological filamentous systems, such as cytoskeleton networks and λ-DNA, and report the dynamics of short actin sliding across a myosin-coated surface. Further, pSIM reveals the side-by-side organization of the actin ring structures in the membrane-associated periodic skeleton of hippocampal neurons and images the dipole dynamics of green fluorescent protein-labeled microtubules in live U2OS cells. pSIM applies directly to a large variety of commercial and home-built SIM systems with various imaging modality.

scholarly journals Customizable live-cell imaging chambers for multimodal and multiplex fluorescence microscopy

2020 ◽  
Vol 98 (5) ◽  
pp. 612-623
Author(s):  
Adam Tepperman ◽  
David Jiao Zheng ◽  
Maria Abou Taka ◽  
Angela Vrieze ◽  
Austin Le Lam ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Super Resolution ◽  
Live Cell ◽  
Imaging Modalities ◽  
Experimental Conditions ◽  
Microscopy Imaging ◽  
Glass Coverslip ◽  
Multiple Imaging ◽  
Microscopy Techniques

Using multiple imaging modalities while performing independent experiments in parallel can greatly enhance the throughput of microscopy-based research, but requires the provision of appropriate experimental conditions in a format that meets the optical requirements of the microscope. Although customized imaging chambers can meet these challenges, the difficulty of manufacturing custom chambers and the relatively high cost and design inflexibility of commercial chambers has limited the adoption of this approach. Herein, we demonstrate the use of 3D printing to produce inexpensive, customized, live-cell imaging chambers that are compatible with a range of imaging modalities, including super-resolution microscopy. In this approach, biocompatible plastics are used to print imaging chambers designed to meet the specific needs of an experiment, followed by adhesion of the printed chamber to a glass coverslip, producing a chamber that is impermeant to liquids and that supports the growth and imaging of cells over multiple days. This approach can also be used to produce moulds for casting microfluidic devices made of polydimethylsiloxane. The utility of these chambers is demonstrated using designs for multiplex microscopy, imaging under shear, chemotaxis, and general cellular imaging. Together, this approach represents an inexpensive yet highly customizable approach for producing imaging chambers that are compatible with modern microscopy techniques.

scholarly journals Fringe optimization for structured illumination super-resolution microscope with digital micromirror device

2019 ◽  
Vol 12 (03) ◽  
pp. 1950014 ◽  
Author(s):  
Xibin Yang ◽  
Qian Zhu ◽  
Zhenglong Sun ◽  
Gang Wen ◽  
Xin Jin ◽  
...  
Keyword(s):  
Modulation Depth ◽  
Low Cost ◽  
Super Resolution ◽  
Fluorescent Labeling ◽  
Live Cells ◽  
Digital Micromirror Device ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Wide Range ◽  
Fringe Patterns

Structured illumination microscopy (SIM) is a promising super-resolution technique for imaging subcellular structures and dynamics due to its compatibility with most commonly used fluorescent labeling methods. Structured illumination can be obtained by either laser interference or projection of fringe patterns. Here, we proposed a fringe projector composed of a compact multi-wavelength LEDs module and a digital micromirror device (DMD) which can be directly attached to most commercial inverted fluorescent microscopes and update it into a SIM system. The effects of the period and duty cycle of fringe patterns on the modulation depth of the structured light field were studied. With the optimized fringe pattern, [Formula: see text] resolution improvement could be obtained with high-end oil objectives. Multicolor imaging and dynamics of subcellular organelles in live cells were also demonstrated. Our method provides a low-cost solution for SIM setup to expand its wide range of applications to most research labs in the field of life science and medicine.

scholarly journals Light-Emitting Diode Based Photoacoustic Imaging System

2020 ◽  
Author(s):  
Ebrahim Najafzadeh ◽  
Parastoo Farnia ◽  
Alireza Ahmadian ◽  
Hossein Ghadiri
Keyword(s):  
Imaging System ◽  
Photoacoustic Imaging ◽  
Light Emitting Diode ◽  
Imaging Modality ◽  
Optical Excitation ◽  
Poly Vinyl Alcohol ◽  
Ultrasound Images ◽  
Light Emitting ◽  
Non Invasive ◽  
Wide Range

Purpose: A Photoacoustic Imaging (PAI) as a non-invasive hybrid imaging modality has the potential to be used in a wide range of pre-clinical and clinical applications. There are different optical excitation sources that affect the performance of PAI systems. Our goal is proving the capability of the Light-Emitting Diode (LED) based PAI system for imaging of objects in different depths. Materials and Methods: In this study the Full Width of Half Maximum (FWHM) and Contrast to Noise Ratio (CNR) of LED-based PAI system is evaluated using agar, and Poly-Vinyl Alcohol Cryogel (PVA-C) phantoms. Results: The results show that axial and lateral FWHM of the photoacoustic image in agar phantom 1%, are 0.59 and 1.16 mm, respectively. It is capable of distinguishing objects about 250 µm. Furthermore, one of the main improvements of photoacoustic images is achieved by proposed LED-based system that is a 26% higher CNR versus the ultrasound images. Conclusion: Therefore, the provided technical characteristics in this study have made designed LED-based PAI system as a suitable tool for preclinical and clinical imaging.

DEEP-UV CONFOCAL FLUORESCENCE IMAGING AND SUPER-RESOLUTION OPTICAL MICROSCOPY OF BIOLOGICAL SAMPLES

2012 ◽  
Vol 05 (04) ◽  
pp. 1250025 ◽  
Author(s):  
TREVOR A. SMITH ◽  
LIISA M. HIRVONEN ◽  
CRAIG N. LINCOLN ◽  
XIAOTAO HAO
Keyword(s):  
Fluorescence Imaging ◽  
Biological Samples ◽  
Super Resolution ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Diffractive Imaging ◽  
Wide Range ◽  
Confocal Fluorescence ◽  
Deep Uv ◽  
Confocal Fluorescence Imaging

A wide range of techniques has been developed to image biological samples at high spatial and temporal resolution. In this paper, we report recent results from deep-UV confocal fluorescence microscopy to image inherent emission from fluorophores such as tryptophan, and structured illumination microscopy (SIM) of biological materials. One motivation for developing deep-UV fluorescence imaging and SIM is to provide methods to complement our measurements in the emerging field of X-ray coherent diffractive imaging.

scholarly journals Customizable Live-Cell Imaging Chambers for Multimodal and Multiplex Fluorescence Microscopy

2020 ◽  
Author(s):  
Adam Tepperman ◽  
David Jiao Zheng ◽  
Maria Abou Taka ◽  
Angela Vrieze ◽  
Austin Le Lam ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Cell Imaging ◽  
Super Resolution ◽  
Live Cell ◽  
Imaging Modalities ◽  
Experimental Conditions ◽  
Microscopy Imaging ◽  
Glass Coverslip ◽  
Multiple Imaging ◽  
Microscopy Techniques

AbstractUsing multiple imaging modalities while performing independent experiments in parallel can greatly enhance the throughput of microscopy-based research, but requires provision of appropriate experimental conditions in a format that meets the microscopy’s optical requirements. Although customized imaging chambers can meet these challenges, the difficulty of manufacturing custom chambers and the relatively high cost and design inflexibility of commercial chambers has limited the adoption of this approach. Herein, we demonstrate the use of 3D printing to produce inexpensive, customized live-cell imaging chambers that are compatible with a range of imaging modalities including super-resolution microscopy. In this approach, biocompatible plastics are used to print imaging chambers designed to meet the specific needs of an experiment, followed by adhesion of the printed chamber to a glass coverslip, producing a chamber that is impermeant to liquids and which supports the growth and imaging of cells over multiple days. This approach can also be used to produce moulds for casting PDMS microfluidic devices. The utility of these chambers is demonstrated using designs for multiplex microscopy, imaging under shear, chemotaxis, and general cellular imaging. Together, this approach represents an inexpensive yet highly customizable approach to produce imaging chambers that are compatible with modern microscopy techniques.

scholarly journals The Valuable Role of Imaging Modalities in the Diagnosis of the Uncommon Presentations of COVID-19: An Educative Case Series

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Mohammadreza Khaleghi ◽  
Alireza Aziz-Ahari ◽  
Nahid Rezaeian ◽  
Sanaz Asadian ◽  
Amirsajjad Mounesi Sohi ◽  
...  
Keyword(s):  
Mesenteric Ischemia ◽  
Imaging Modality ◽  
Brain Mri ◽  
Clinical Signs ◽  
Case Series ◽  
Imaging Modalities ◽  
Adequate Treatment ◽  
Abdominal Symptoms ◽  
First Case ◽  
Wide Range

The outbreak of coronavirus disease 2019 (COVID-19) in late 2019 rapidly turned into a global pandemic. Although the symptoms of COVID-19 are mainly respiratory ones, the infection is associated with a wide range of clinical signs and symptoms. The main imaging modality in COVID-19 is lung computed tomography (CT) scanning, but the diagnosis of the vast spectrum of complications needs the application of various imaging modalities. Owing to the novelty of the disease and its presentations, its complications—particularly uncommon ones—can be easily missed. In this study, we describe some uncommon presentations of COVID-19 diagnosed by various imaging modalities. The first case presented herein was a man with respiratory distress, who transpired to suffer from pneumothorax and pneumomediastinum in addition to the usual pneumonia of COVID-19. The second patient was a hospitalized COVID-19 case, whose clinical condition suddenly deteriorated with the development of abdominal symptoms diagnosed as mesenteric ischemia by abdominal CT angiography. The third patient was a case of cardiac involvement in the COVID-19 course, detected as myocarditis by cardiac magnetic resonance imaging (MRI). The fourth and fifth cases were COVID-19-associated encephalitis whose diagnoses were established by brain MRI. COVID-19 is a multisystem disorder with a wide range of complications such as pneumothorax, pneumomediastinum, mesenteric ischemia, myocarditis, and encephalitis. Prompt diagnosis with appropriate imaging modalities can lead to adequate treatment and better survival.

scholarly journals Super-resolution structured illumination microscopy: past, present and future

2021 ◽  
Vol 379 (2199) ◽  
Author(s):  
Kirti Prakash ◽  
Benedict Diederich ◽  
Stefanie Reichelt ◽  
Rainer Heintzmann ◽  
Lothar Schermelleh
Keyword(s):  
Three Dimensional ◽  
Super Resolution ◽  
Computational Imaging ◽  
Biological Applications ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Recent Developments ◽  
Resolution Imaging ◽  
Microscopy Techniques ◽  
Super Resolution Microscopy

Structured illumination microscopy (SIM) has emerged as an essential technique for three-dimensional (3D) and live-cell super-resolution imaging. However, to date, there has not been a dedicated workshop or journal issue covering the various aspects of SIM, from bespoke hardware and software development and the use of commercial instruments to biological applications. This special issue aims to recap recent developments as well as outline future trends. In addition to SIM, we cover related topics such as complementary super-resolution microscopy techniques, computational imaging, visualization and image processing methods.This article is part of the Theo Murphy meeting issue ‘Super-resolution structured illumination microscopy (part 1)’.

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