Reconstruction in wide-field interferometric microscopy for imaging weakly scattering biological nanoparticles with super-resolution (Conference Presentation)

2018 ◽  
Author(s):  
Oguzhan Avci ◽  
Celalettin Yurdakul ◽  
Derin D. Sevenler ◽  
Fulya E. Kanik ◽  
Alex C. Matlock ◽  
...  
Keyword(s):  
Super Resolution ◽  
Wide Field

scholarly journals Comparing Super-Resolution Microscopy Techniques to Analyze Chromosomes

2021 ◽  
Vol 22 (4) ◽  
pp. 1903
Author(s):  
Ivona Kubalová ◽  
Alžběta Němečková ◽  
Klaus Weisshart ◽  
Eva Hřibová ◽  
Veit Schubert
Keyword(s):  
Single Molecule ◽  
Imaging Techniques ◽  
Chromatin Condensation ◽  
Super Resolution ◽  
Stimulated Emission ◽  
Wide Field ◽  
Structured Illumination Microscopy ◽  
Metaphase Chromosomes ◽  
Localization Microscopy ◽  
Super Resolution Microscopy

The importance of fluorescence light microscopy for understanding cellular and sub-cellular structures and functions is undeniable. However, the resolution is limited by light diffraction (~200–250 nm laterally, ~500–700 nm axially). Meanwhile, super-resolution microscopy, such as structured illumination microscopy (SIM), is being applied more and more to overcome this restriction. Instead, super-resolution by stimulated emission depletion (STED) microscopy achieving a resolution of ~50 nm laterally and ~130 nm axially has not yet frequently been applied in plant cell research due to the required specific sample preparation and stable dye staining. Single-molecule localization microscopy (SMLM) including photoactivated localization microscopy (PALM) has not yet been widely used, although this nanoscopic technique allows even the detection of single molecules. In this study, we compared protein imaging within metaphase chromosomes of barley via conventional wide-field and confocal microscopy, and the sub-diffraction methods SIM, STED, and SMLM. The chromosomes were labeled by DAPI (4′,6-diamidino-2-phenylindol), a DNA-specific dye, and with antibodies against topoisomerase IIα (Topo II), a protein important for correct chromatin condensation. Compared to the diffraction-limited methods, the combination of the three different super-resolution imaging techniques delivered tremendous additional insights into the plant chromosome architecture through the achieved increased resolution.

scholarly journals Camera-based localization microscopy optimized with calibrated structured illumination

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Martin Schmidt ◽  
Adam C. Hundahl ◽  
Henrik Flyvbjerg ◽  
Rodolphe Marie ◽  
Kim I. Mortensen
Keyword(s):  
Data Analysis ◽  
Super Resolution ◽  
Wide Field ◽  
Structured Illumination ◽  
Uniform Illumination ◽  
Localization Microscopy ◽  
Localization And Tracking ◽  
Field Imaging ◽  
Wide Field Imaging ◽  
Localization Information

AbstractUntil very recently, super-resolution localization and tracking of fluorescent particles used camera-based wide-field imaging with uniform illumination. Then it was demonstrated that structured illuminations encode additional localization information in images. The first demonstration of this uses scanning and hence suffers from limited throughput. This limitation was mitigated by fusing camera-based localization with wide-field structured illumination. Current implementations, however, use effectively only half the localization information that they encode in images. Here we demonstrate how all of this information may be exploited by careful calibration of the structured illumination. Our approach achieves maximal resolution for given structured illumination, has a simple data analysis, and applies to any structured illumination in principle. We demonstrate this with an only slightly modified wide-field microscope. Our protocol should boost the emerging field of high-precision localization with structured illumination.

scholarly journals A technique for resolution assessment in blind-SIM experiments

2021 ◽  
Author(s):  
Imen Boujmil ◽  
Giancarlo Ruocco ◽  
Marco Leonetti
Keyword(s):  
High Resolution ◽  
Biological Sample ◽  
A Priori ◽  
Resolution Enhancement ◽  
Super Resolution ◽  
Experimental Setup ◽  
Wide Field ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Resolution Measurement

Super resolution techniques are an excellent alternative to wide field microscopy, providing high resolution also in (typically fragile) biological sample. Among the various super resolution techniques, Structured Illumination Microscopy (SIM) improve resolution by employing multiple illumination patterns to be deconvolved with a dedicated software. In the case of blind SIM techniques, unknown patterns, such as speckles, are used, thus providing super resolved images, nearly unaffected by aberrations with a simplified experimental setup. Scattering Assisted Imaging, a special blind SIM technique, exploits an illumination PSF (speckle grains size), smaller than the collection PSF (defined by the collection objectives), to surpass the typical SIM resolution enhancement. However, if SAI is used, it is very difficult to extract the resolution enhancement form a priori considerations. In this paper we propose a protocol and experimental setup for the resolution measurement, demonstrating the resolution enhancement for different collection PSF values.

scholarly journals ELM: super-resolution analysis of wide-field images of fluorescent shell structures

2018 ◽  
Vol 6 (3) ◽  
pp. 037001 ◽  
Author(s):  
James D Manton ◽  
Yao Xiao ◽  
Robert D Turner ◽  
Graham Christie ◽  
Eric J Rees
Keyword(s):  
Super Resolution ◽  
Shell Structures ◽  
Wide Field ◽  
Resolution Analysis

Applications of nanostructures in wide-field, label-free super resolution microscopy

2018 ◽  
Vol 27 (11) ◽  
pp. 118704 ◽  
Author(s):  
Xiaowei Liu ◽  
Chao Meng ◽  
Xuechu Xu ◽  
Mingwei Tang ◽  
Chenlei Pang ◽  
...  
Keyword(s):  
Super Resolution ◽  
Wide Field ◽  
Label Free ◽  
Super Resolution Microscopy

scholarly journals An Easy Path for Correlative Electron and Super-Resolution Light Microscopy

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Dorothea Pinotsi ◽  
Simona Rodighiero ◽  
Silvia Campioni ◽  
Gabor Csucs
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Light Microscopy ◽  
Amyloid Fibrils ◽  
Light And Electron Microscopy ◽  
Super Resolution ◽  
Wide Field ◽  
Set Up ◽  
Bio Compatibility ◽  
Scanning Electron

Abstract A number of new Correlative Light and Electron Microscopy approaches have been developed over the past years, offering the opportunity to combine the specificity and bio-compatibility of light microscopy with the high resolution achieved in electron microscopy. More recently, these approaches have taken one step further and also super-resolution light microscopy was combined with transmission or scanning electron microscopy. This combination usually requires moving the specimen between different imaging systems, an expensive set-up and relatively complicated imaging workflows. Here we present a way to overcome these difficulties by exploiting a commercially available wide-field fluorescence microscope integrated in the specimen chamber of a Scanning Electron Microscope (SEM) to perform correlative LM/EM studies. Super-resolution light microscopy was achieved by using a recently developed algorithm - the Super-Resolution Radial Fluctuations (SRRF) - to improve the resolution of diffraction limited fluorescent images. With this combination of hardware/software it is possible to obtain correlative super-resolution light and scanning electron microscopy images in an easy and fast way. The imaging workflow is described and demonstrated on fluorescently labelled amyloid fibrils, fibrillar protein aggregates linked to the onset of multiple neurodegenerative diseases, revealing information about their polymorphism.

Mapping Pixel Dissimilarity in Wide-Field Super-Resolution Fluorescence Microscopy

2015 ◽  
Vol 87 (9) ◽  
pp. 4675-4682 ◽  
Author(s):  
Cyril Ruckebusch ◽  
Romain Bernex ◽  
Franco Allegrini ◽  
Michel Sliwa ◽  
Johan Hofkens ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Super Resolution ◽  
Wide Field

scholarly journals Super-resolution endoscopy for real-time wide-field imaging

2015 ◽  
Vol 23 (13) ◽  
pp. 16803 ◽  
Author(s):  
Feifei Wang ◽  
Hok Sum Sam Lai ◽  
Lianqing Liu ◽  
Pan Li ◽  
Haibo Yu ◽  
...  
Keyword(s):  
Real Time ◽  
Super Resolution ◽  
Wide Field ◽  
Field Imaging ◽  
Wide Field Imaging

scholarly journals Quantitative comparison of camera technologies for cost-effective Super-resolution Optical Fluctuation Imaging (SOFI)

2018 ◽  
Author(s):  
Robin Van den Eynde ◽  
Alice Sandmeyer ◽  
Wim Vandenberg ◽  
Sam Duwé ◽  
Wolfgang Hübner ◽  
...  
Keyword(s):  
Live Cell Imaging ◽  
Imaging System ◽  
Fluorescent Proteins ◽  
Imaging Techniques ◽  
Super Resolution ◽  
Cost Effective ◽  
Wide Field ◽  
Cmos Camera ◽  
Resolution Imaging ◽  
Cost Efficient

AbstractSuper-Resolution (SR) fluorescence microscopy is typically carried out on high-end research microscopes. Super-resolution Optical Fluctuation Imaging (SOFI) is a fast SR technique capable of live-cell imaging, that is compatible with many wide-field microscope systems. However, especially when employing fluorescent proteins, a key part of the imaging system is a very sensitive and well calibrated camera sensor. The substantial costs of such systems preclude many research groups from employing super-resolution imaging techniques.Here, we examine to what extent SOFI can be performed using a range of imaging hardware comprising different technologies and costs. In particular, we quantitatively compare the performance of an industry-grade CMOS camera to both state-of-the-art emCCD and sCMOS detectors, with SOFI-specific metrics. We show that SOFI data can be obtained using a cost-efficient industry-grade sensor, both on commercial and home-built microscope systems, though our analysis also readily exposes the merits of the per-pixel corrections performed in scientific cameras.

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