scholarly journals Quantitative Analysis of Interactive Behavior of Mitochondria and Lysosomes Using Structured Illumination Microscopy

2018 ◽  
Author(s):  
Qixin Chen ◽  
Xintian Shao ◽  
Mingang Hao ◽  
Zhiqi Tian ◽  
Chenran Wang ◽  
...  
Keyword(s):  
Quantitative Analysis ◽  
Optical Microscopy ◽  
Spatial Resolution ◽  
Cell Biology ◽  
Super Resolution ◽  
Cellular Level ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Interactive Behavior ◽  
Molecular Machinery

ABSTRACTSuper-resolution optical microscopy has extended the spatial resolution of cell biology from the cellular level to the nanoscale, enabling the observation of the interactive behavior of single mitochondria and lysosomes. Quantitative parametrization of interaction between mitochondria and lysosomes under super-resolution optical microscopy, however, is currently unavailable, which has severely limited our understanding of the molecular machinery underlying mitochondrial functionality. Here, we introduce an M-value to quantitatively investigate mitochondria and lysosome contact (MLC) and mitophagy under structured illumination microscopy. We found that the M-value for an MLC is typically less than 0.4, whereas in mitophagy it ranges from 0.5 to 1.0. This system permits further investigation of the detailed molecular mechanism governing the interactive behavior of mitochondria and lysosomes.

scholarly journals Structured illumination microscopy and its new developments

2016 ◽  
Vol 09 (03) ◽  
pp. 1630010 ◽  
Author(s):  
Jianling Chen ◽  
Caimin Qiu ◽  
Minghai You ◽  
Xiaogang Chen ◽  
Hongqin Yang ◽  
...  
Keyword(s):  
Optical Microscopy ◽  
Spatial Resolution ◽  
Imaging Techniques ◽  
Super Resolution ◽  
Living Cells ◽  
The Other ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Recent Developments ◽  
Super Resolution Microscopy

Optical microscopy allows us to observe the biological structures and processes within living cells. However, the spatial resolution of the optical microscopy is limited to about half of the wavelength by the light diffraction. Structured illumination microscopy (SIM), a type of new emerging super-resolution microscopy, doubles the spatial resolution by illuminating the specimen with a patterned light, and the sample and light source requirements of SIM are not as strict as the other super-resolution microscopy. In addition, SIM is easier to combine with the other imaging techniques to improve their imaging resolution, leading to the developments of diverse types of SIM. SIM has great potential to meet the various requirements of living cells imaging. Here, we review the recent developments of SIM and its combination with other imaging techniques.

scholarly journals Quantitative analysis of interactive behavior of mitochondria and lysosomes using structured illumination microscopy

Biomaterials ◽  
2020 ◽  
Vol 250 ◽  
pp. 120059 ◽  
Author(s):  
Qixin Chen ◽  
Xintian Shao ◽  
Mingang Hao ◽  
Hongbao Fang ◽  
Ruilin Guan ◽  
...  
Keyword(s):  
Quantitative Analysis ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Interactive Behavior

scholarly journals Visualizing ultrastructural details of placental tissue with super-resolution structured illumination microscopy

2020 ◽  
Author(s):  
Luis E. Villegas-Hernández ◽  
Mona Nystad ◽  
Florian Ströhl ◽  
Purusotam Basnet ◽  
Ganesh Acharya ◽  
...  
Keyword(s):  
Cell Biology ◽  
Resolution Enhancement ◽  
Super Resolution ◽  
Placental Tissue ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Chorionic Villi ◽  
Tissue Sections ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded

AbstractSuper-resolution fluorescence microscopy is a widely employed technique in cell biology research, yet remains relatively unexplored in the field of histo-pathology. Here, we describe the sample preparation steps and acquisition parameters necessary to obtain fluorescent multicolor super-resolution structured illumination microscopy (SIM) images of both formalin-fixed paraffin-embedded and cryo-preserved placental tissue sections. We compare super-resolved images of chorionic villi against diffraction-limited deconvolution microscopy and demonstrate the significant contrast and resolution enhancement attainable with SIM. We show that SIM resolves ultrastructural details such as the syncytiotrophoblast’s microvilli brush border, which up until now has been only resolvable by electron microscopy.

scholarly journals Simultaneous Zn2+ tracking in multiple organelles using super-resolution morphology-correlated organelle identification in living cells

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Hongbao Fang ◽  
Shanshan Geng ◽  
Mingang Hao ◽  
Qixin Chen ◽  
Minglun Liu ◽  
...  
Keyword(s):  
Stem Cell ◽  
Fluorescent Probe ◽  
Cell Biology ◽  
Pluripotent Stem Cell ◽  
Reliable Method ◽  
Super Resolution ◽  
Living Cells ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Turn On

AbstractZn2+ plays important roles in metabolism and signaling regulation. Subcellular Zn2+ compartmentalization is essential for organelle functions and cell biology, but there is currently no method to determine Zn2+ signaling relationships among more than two different organelles with one probe. Here, we report simultaneous Zn2+ tracking in multiple organelles (Zn-STIMO), a method that uses structured illumination microscopy (SIM) and a single Zn2+ fluorescent probe, allowing super-resolution morphology-correlated organelle identification in living cells. To guarantee SIM imaging quality for organelle identification, we develop a new turn-on Zn2+ fluorescent probe, NapBu-BPEA, by regulating the lipophilicity of naphthalimide-derived Zn2+ probes to make it accumulate in multiple organelles except the nucleus. Zn-STIMO with this probe shows that CCCP-induced mitophagy in HeLa cells is associated with labile Zn2+ enhancement. Therefore, direct organelle identification supported by SIM imaging makes Zn-STIMO a reliable method to determine labile Zn2+ dynamics in various organelles with one probe. Finally, SIM imaging of pluripotent stem cell-derived organoids with NapBu-BPEA demonstrates the potential of super-resolution morphology-correlated organelle identification to track biospecies and events in specific organelles within organoids.

scholarly journals Improving axial resolution in Structured Illumination Microscopy using deep learning

2021 ◽  
Vol 379 (2199) ◽  
Author(s):  
Miguel A. Boland ◽  
Edward A. K. Cohen ◽  
Seth R. Flaxman ◽  
Mark A. A. Neil
Keyword(s):  
Deep Learning ◽  
Optical Microscopy ◽  
Super Resolution ◽  
Structured Illumination ◽  
Learning Models ◽  
Axial Resolution ◽  
Structured Illumination Microscopy ◽  
Recent Advances ◽  
Robust To Noise ◽  
Up Scaling

Structured Illumination Microscopy (SIM) is a widespread methodology to image live and fixed biological structures smaller than the diffraction limits of conventional optical microscopy. Using recent advances in image up-scaling through deep learning models, we demonstrate a method to reconstruct 3D SIM image stacks with twice the axial resolution attainable through conventional SIM reconstructions. We further demonstrate our method is robust to noise and evaluate it against two-point cases and axial gratings. Finally, we discuss potential adaptions of the method to further improve resolution. This article is part of the Theo Murphy meeting issue ‘Super-resolution structured illumination microscopy (part 1)’.

Turn-key super-resolution mapping of cell receptor force orientation and magnitude using a commercial structured illumination microscope

2020 ◽  
Author(s):  
Aaron Blanchard ◽  
J. Dale Combs ◽  
Joshua Brockman ◽  
Anna Kellner ◽  
Roxanne Glazier ◽  
...  
Keyword(s):  
Cell Biology ◽  
Focal Adhesions ◽  
Super Resolution ◽  
Cell Receptor ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Surface Receptors ◽  
Cellular Processes ◽  
Resolution Mapping ◽  
Super Resolution Mapping

Abstract Many cellular processes, including cell division, development, and cell migration require spatially and temporally coordinated forces transduced by cell surface receptors. Nucleic acid-based molecular tension probes allow one to quantify and visualize the piconewton (pN) forces applied by these receptors. Building on this technology, we recently imaged DNA tension probes using fluorescence polarization imaging to map the magnitude and 3D orientation of receptor forces with diffraction limited resolution (~ 250 nm). Further improvements in spatial resolution are desirable as many force-sensing receptors are organized at the nano-scale in supramolecular complexes such as focal adhesions. Here, we show that structured illumination microscopy (SIM), a super-resolution technique, can be used to perform super-resolution molecular force microscopy (MFM). Using SIM-MFM, we generate the highest resolution maps of both the magnitude and orientation of the pN traction forces applied by cells. We apply SIM-MFM to map platelet and fibroblast integrins forces, as well as T cell receptor forces. The method reveals that platelets dynamically re-arrange the orientation of their integrin forces during activation. Monte Carlo simulations validated the results and provided analysis of the sources of noise. Importantly, we envision that SIM-MFM will be broadly adopted by the cell biology and mechanobiology communities because it can be implemented on any standard SIM microscope without hardware modifications.

scholarly journals SIMcheck: a Toolbox for Successful Super-resolution Structured Illumination Microscopy

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Graeme Ball ◽  
Justin Demmerle ◽  
Rainer Kaufmann ◽  
Ilan Davis ◽  
Ian M. Dobbie ◽  
...  
Keyword(s):  
Cell Biology ◽  
Three Dimensional ◽  
Super Resolution ◽  
Quality Data ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
High Quality Data ◽  
Accessible Method ◽  
Objective Control ◽  
Common Problems

Abstract Three-dimensional structured illumination microscopy (3D-SIM) is a versatile and accessible method for super-resolution fluorescence imaging, but generating high-quality data is challenging, particularly for non-specialist users. We present SIMcheck, a suite of ImageJ plugins enabling users to identify and avoid common problems with 3D-SIM data and assess resolution and data quality through objective control parameters. Additionally, SIMcheck provides advanced calibration tools and utilities for common image processing tasks. This open-source software is applicable to all commercial and custom platforms and will promote routine application of super-resolution SIM imaging in cell biology.

scholarly journals Structured illumination microscopy and image scanning microscopy: a review and comparison of imaging properties

2021 ◽  
Vol 379 (2199) ◽  
Author(s):  
Colin J. R. Sheppard
Keyword(s):  
Spatial Resolution ◽  
Super Resolution ◽  
Scanning Microscopy ◽  
Structured Illumination ◽  
Practical Application ◽  
Structured Illumination Microscopy ◽  
Image Scanning ◽  
Imaging Performance ◽  
Imaging Properties

Structured illumination microscopy and image scanning microscopy are two microscopical tech- niques, rapidly increasing in practical application, that can result in improvement in transverse spatial resolution, and/or improvement in axial imaging performance. The history and principles of these techniques are reviewed, and the imaging properties of the two methods compared. This article is part of the Theo Murphy meeting issue ‘Super-resolution structured illumination microscopy (part 1)’.

scholarly journals New ways of looking at very small holes – using optical nanoscopy to visualize liver sinusoidal endothelial cell fenestrations

Nanophotonics ◽  
2018 ◽  
Vol 7 (3) ◽  
pp. 575-596 ◽  
Author(s):  
Cristina I. Øie ◽  
Viola Mönkemöller ◽  
Wolfgang Hübner ◽  
Mark Schüttpelz ◽  
Hong Mao ◽  
...  
Keyword(s):  
Single Molecule ◽  
Cell Biology ◽  
Super Resolution ◽  
Living Cells ◽  
Liver Sinusoidal Endothelial Cell ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Liver Sinusoidal Endothelial Cells ◽  
Dynamic Structures

AbstractSuper-resolution fluorescence microscopy, also known as nanoscopy, has provided us with a glimpse of future impacts on cell biology. Far-field optical nanoscopy allows, for the first time, the study of sub-cellular nanoscale biological structures in living cells, which in the past was limited to electron microscopy (EM) (in fixed/dehydrated) cells or tissues. Nanoscopy has particular utility in the study of “fenestrations” – phospholipid transmembrane nanopores of 50–150 nm in diameter through liver sinusoidal endothelial cells (LSECs) that facilitate the passage of plasma, but (usually) not blood cells, to and from the surrounding hepatocytes. Previously, these fenestrations were only discernible with EM, but now they can be visualized in fixed and living cells using structured illumination microscopy (SIM) and in fixed cells using single molecule localization microscopy (SMLM) techniques such as direct stochastic optical reconstruction microscopy. Importantly, both methods use wet samples, avoiding dehydration artifacts. The use of nanoscopy can be extended to the in vitro study of fenestration dynamics, to address questions such as the following: are they actually dynamic structures, and how do they respond to endogenous and exogenous agents? A logical further extension of these methodologies to liver research (including the liver endothelium) will be their application to liver tissue sections from animal models with different pathological manifestations and ultimately to patient biopsies. This review will cover the current state of the art of the use of nanoscopy in the study of liver endothelium and the liver in general. Potential future applications in cell biology and the clinical implications will be discussed.

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