Special Issue "Superresolution Optical Microscopy"

A special issue of Photonics (ISSN 2304-6732).

Deadline for manuscript submissions: closed (30 April 2017)

Special Issue Editors

Guest Editor
Prof. Dr. Christoph Cremer

1. Superresolution Microscopy, Institute of Molecular Biology (IMB), D-55128 Mainz, Germany
2. Kirchhoff Institute of Physics (KIP), University Heidelberg, Germany
Website | E-Mail
Interests: development and application of methods of super-resolving fluorescence light microscopy: Focused nanoscopy, structured illumination, various types of localization microscopy
Guest Editor
Prof. Dr. Rainer Heintzmann

1. Nanobiophotonics Professor for Physical Chemistry, Institute of Physical Chemistry, Friedrich-Schiller-Universität Jena, Helmholtzweg 4, 07743 Jena, Germany
2. Head of the Microscopy Reseach Unit, Leibniz Institute of Photonic Technology, Albert-Einstein Str. 9, 07745 Jena, Germany
Website | E-Mail
Interests: high resolution fluorescence microscopy, image processing, deconvolution algorithms, optical coherence microscopy, holography, Integrated field spectroscopy, hyperspectral Raman imaging
Guest Editor
Prof. Dr. Jennifer Lippincott-Schwartz

Howard Hughes Medical Institute - Janelia Research Campus 19700 Helix Drive Ashburn, VA 20147
Website | E-Mail
Phone: 571-209-4133

Special Issue Information

Dear Colleagues,

Presently, a variety of methods exist that overcome the hundred-year-old theoretical limit for the resolution potential of light microscopy (of about 200 nm for visible light), which for many decades has precluded a direct glimpse of the molecular machinery of life. In this Special Issue, we plan to summarize the state of the art from the methodological point of view, and to address some of the challenges still to be resolved before the SRM approaches will be fit to bring about the revolution envisaged in biology, medicine, and the material sciences. In this context, contributions to these subjects are welcome to include optical, physico-chemical, technical and image processing topics.

Prof. Dr. Christoph Cremer
Prof. Dr. Rainer Heintzmann
Prof. Dr. Jennifer Lippincott-Schwartz
Guest Editor

Manuscript Submission Information

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Keywords

  • Methods of SRM (Superresolution microscopy), e.g. focused nanoscopy like 4Pi, STED, RESOLFT; localization microscopy; structured illumination; expansion microscopy
  • Resolution issues, e.g. localization precision & accuracy, labelling efficiency, optical resolution, structural resolution, emitter density, Nyquist and Fourier ring correlated criteria
  • Photophysics of SRM, e.g. light induced photoconversation, “bright-dark” and “dark-bright” transitions; use of photostable emitters
  • Labelling techniques, e.g. fluorescent proteins, standard dyes, special dyes
  • Detection technology, image processing, deconvolution, data mining, data storage
  • Applications of SRM (e.g. membranes, nuclear nanostructure, bacteria, viruses; material sciences)
  • Perspectives of SRM (e.g. in terms of instrumentation, resolution, in vivo, high throughput, working distance, field of view)
  •  Sample preparation techniques and protocols specifically relevant to SRM.

Published Papers (7 papers)

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Research

Open AccessArticle Exploring the Potential of Airyscan Microscopy for Live Cell Imaging
Received: 1 June 2017 / Revised: 19 June 2017 / Accepted: 30 June 2017 / Published: 7 July 2017
Cited by 7 | PDF Full-text (3570 KB) | HTML Full-text | XML Full-text
Abstract
Biological research increasingly demands the use of non-invasive and ultra-sensitive imaging techniques. The Airyscan technology was recently developed to bridge the gap between conventional confocal and super-resolution microscopy. This technique combines confocal imaging with a 0.2 Airy Unit pinhole, deconvolution and the pixel-reassignment
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Biological research increasingly demands the use of non-invasive and ultra-sensitive imaging techniques. The Airyscan technology was recently developed to bridge the gap between conventional confocal and super-resolution microscopy. This technique combines confocal imaging with a 0.2 Airy Unit pinhole, deconvolution and the pixel-reassignment principle in order to enhance both the spatial resolution and signal-to-noise-ratio without increasing the excitation power and acquisition time. Here, we present a detailed study evaluating the performance of Airyscan as compared to confocal microscopy by imaging a variety of reference samples and biological specimens with different acquisition and processing parameters. We found that the processed Airyscan images at default deconvolution settings have a spatial resolution similar to that of conventional confocal imaging with a pinhole setting of 0.2 Airy Units, but with a significantly improved signal-to-noise-ratio. Further gains in the spatial resolution could be achieved by the use of enhanced deconvolution filter settings, but at a steady loss in the signal-to-noise ratio, which at more extreme settings resulted in significant data loss and image distortion. Full article
(This article belongs to the Special Issue Superresolution Optical Microscopy)
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Open AccessArticle Mapping Molecular Function to Biological Nanostructure: Combining Structured Illumination Microscopy with Fluorescence Lifetime Imaging (SIM + FLIM)
Received: 16 May 2017 / Revised: 30 June 2017 / Accepted: 30 June 2017 / Published: 7 July 2017
Cited by 2 | PDF Full-text (1556 KB) | HTML Full-text | XML Full-text | Supplementary Files
Abstract
We present a new microscope integrating super-resolved imaging using structured illumination microscopy (SIM) with wide-field optically sectioned fluorescence lifetime imaging (FLIM) to provide optical mapping of molecular function and its correlation with biological nanostructure below the conventional diffraction limit. We illustrate this SIM
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We present a new microscope integrating super-resolved imaging using structured illumination microscopy (SIM) with wide-field optically sectioned fluorescence lifetime imaging (FLIM) to provide optical mapping of molecular function and its correlation with biological nanostructure below the conventional diffraction limit. We illustrate this SIM + FLIM capability to map FRET readouts applied to the aggregation of discoidin domain receptor 1 (DDR1) in Cos 7 cells following ligand stimulation and to the compaction of DNA during the cell cycle. Full article
(This article belongs to the Special Issue Superresolution Optical Microscopy)
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Open AccessArticle Phase Mask-Based Multimodal Superresolution Microscopy
Received: 2 May 2017 / Revised: 22 June 2017 / Accepted: 30 June 2017 / Published: 6 July 2017
Cited by 2 | PDF Full-text (1127 KB) | HTML Full-text | XML Full-text
Abstract
We demonstrate a multimodal superresolution microscopy technique based on a phase masked excitation beam in combination with spatially filtered detection. The theoretical foundation for calculating the focus from a non-paraxial beam with an arbitrary azimuthally symmetric phase mask is presented for linear and
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We demonstrate a multimodal superresolution microscopy technique based on a phase masked excitation beam in combination with spatially filtered detection. The theoretical foundation for calculating the focus from a non-paraxial beam with an arbitrary azimuthally symmetric phase mask is presented for linear and two-photon excitation processes as well as the theoretical resolution limitations. Experimentally this technique is demonstrated using two-photon luminescence from 80 nm gold particle as well as two-photon fluorescence lifetime imaging of fluorescent polystyrene beads. Finally to illustrate the versatility of this technique we acquire two-photon fluorescence lifetime, two-photon luminescence, and second harmonic images of a mixture of fluorescent molecules and 80 nm gold particles with <120 nm resolution ( λ /7). Since this approach exclusively relies on engineering the excitation and collection volumes, it is suitable for a wide range of scanning-based microscopies. Full article
(This article belongs to the Special Issue Superresolution Optical Microscopy)
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Open AccessArticle Analytical Model of the Optical Vortex Scanning Microscope with a Simple Phase Object
Received: 14 March 2017 / Revised: 20 May 2017 / Accepted: 30 May 2017 / Published: 5 June 2017
Cited by 4 | PDF Full-text (3986 KB) | HTML Full-text | XML Full-text
Abstract
An analytical model of an optical vortex microscope, in which a simple phase object was inserted into the illuminating beam, is presented. In this microscope, the focused vortex beam interacts with an object and transmits the corresponding information to the detection plane. It
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An analytical model of an optical vortex microscope, in which a simple phase object was inserted into the illuminating beam, is presented. In this microscope, the focused vortex beam interacts with an object and transmits the corresponding information to the detection plane. It was shown that the beam at the detection plane can be separated analytically into two parts: a non-disturbed vortex part and an object beam part. The intensity of the non-disturbed part spreads out over the center; hence, the small disturbance introduced by the object can be detected at the image center. A first procedure for recovering information about the object from this set-up was proposed. The theory was verified experimentally. Full article
(This article belongs to the Special Issue Superresolution Optical Microscopy)
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Open AccessArticle Label-Free Saturated Structured Excitation Microscopy
Received: 14 March 2017 / Revised: 17 April 2017 / Accepted: 2 May 2017 / Published: 5 May 2017
PDF Full-text (2220 KB) | HTML Full-text | XML Full-text
Abstract
Micro- and nanoscale chemical and structural heterogeneities, whether they are intrinsic material properties like grain boundaries or intentionally encoded via nanoscale fabrication techniques, pose a challenge to current material characterization methods. To precisely interrogate the electronic structure of these complex materials systems, spectroscopic
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Micro- and nanoscale chemical and structural heterogeneities, whether they are intrinsic material properties like grain boundaries or intentionally encoded via nanoscale fabrication techniques, pose a challenge to current material characterization methods. To precisely interrogate the electronic structure of these complex materials systems, spectroscopic techniques with high spatial resolution are required. However, conventional optical microscopies are limited to probe volumes of ~200 nm due to the diffraction limit of visible light. While a variety of sub-diffraction-limited techniques have been developed, many rely on fluorescent contrast agents. Herein we describe label-free saturated structured excitation microscopy (LF-SSEM) applicable to nonlinear imaging approaches such as stimulated Raman and pump-probe microscopy. By exploiting the nonlinear sample response of saturated excitation, LF-SSEM provides theoretically limitless resolution enhancement without the need for a photoluminescent sample. Full article
(This article belongs to the Special Issue Superresolution Optical Microscopy)
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Open AccessArticle XL-SIM: Extending Superresolution into Deeper Layers
Received: 14 March 2017 / Revised: 7 April 2017 / Accepted: 13 April 2017 / Published: 20 April 2017
Cited by 2 | PDF Full-text (6471 KB) | HTML Full-text | XML Full-text | Supplementary Files
Abstract
Of all 3D-super resolution techniques, structured illumination microscopy (SIM) provides the best compromise with respect to resolution, signal-to-noise ratio (S/N), speed and cell viability. Its ability to achieve double resolution in all three dimensions enables resolving 3D-volumes almost 10× smaller than with a
[...] Read more.
Of all 3D-super resolution techniques, structured illumination microscopy (SIM) provides the best compromise with respect to resolution, signal-to-noise ratio (S/N), speed and cell viability. Its ability to achieve double resolution in all three dimensions enables resolving 3D-volumes almost 10× smaller than with a normal light microscope. Its major drawback is noise contained in the out-of-focus-signal, which—unlike the out-of-focus signal itself—cannot be removed mathematically. The resulting “noise-pollution” grows bigger the more light is removed, thus rendering thicker biological samples unsuitable for SIM. By using a slit confocal pattern, we employ optical means to suppress out-of-focus light before its noise can spoil SIM mathematics. This not only increases tissue penetration considerably, but also provides a better S/N performance and an improved confocality. The SIM pattern we employ is no line grid, but a two-dimensional hexagonal structure, which makes pattern rotation between image acquisitions obsolete and thus simplifies image acquisition and yields more robust fit parameters for SIM. Full article
(This article belongs to the Special Issue Superresolution Optical Microscopy)
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Open AccessArticle Photothermal Microscopy for High Sensitivity and High Resolution Absorption Contrast Imaging of Biological Tissues
Received: 13 March 2017 / Revised: 4 April 2017 / Accepted: 13 April 2017 / Published: 19 April 2017
Cited by 2 | PDF Full-text (3211 KB) | HTML Full-text | XML Full-text
Abstract
Photothermal microscopy is useful to visualize the distribution of non-fluorescence chromoproteins in biological specimens. Here, we developed a high sensitivity and high resolution photothermal microscopy with low-cost and compact laser diodes as light sources. A new detection scheme for improving signal to noise
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Photothermal microscopy is useful to visualize the distribution of non-fluorescence chromoproteins in biological specimens. Here, we developed a high sensitivity and high resolution photothermal microscopy with low-cost and compact laser diodes as light sources. A new detection scheme for improving signal to noise ratio more than 4-fold is presented. It is demonstrated that spatial resolution in photothermal microscopy is up to nearly twice as high as that in the conventional widefield microscopy. Furthermore, we demonstrated the ability for distinguishing or identifying biological molecules with simultaneous muti-wavelength imaging. Simultaneous photothermal and fluorescence imaging of mouse brain tissue was conducted to visualize both neurons expressing yellow fluorescent protein and endogenous non-fluorescent chromophores. Full article
(This article belongs to the Special Issue Superresolution Optical Microscopy)
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