scholarly journals Structured illumination microscopy for dual-modality 3D sub-diffraction resolution fluorescence and refractive-index reconstruction

2017 ◽  
Vol 8 (12) ◽  
pp. 5776 ◽  
Author(s):  
Shwetadwip Chowdhury ◽  
Will J. Eldridge ◽  
Adam Wax ◽  
Joseph A. Izatt
2021 ◽  
Author(s):  
Yanquan Mo ◽  
Fan Feng ◽  
Heng Mao ◽  
Junchao Fan ◽  
Liangyi Chen

AbstractDespite its wide application in live-cell super-resolution (SR) imaging, structured illumination microscopy (SIM) suffers from aberrations caused by various sources. Although artifacts generated from inaccurate reconstruction parameter estimation and noise amplification can be minimized, aberrations due to the scattering of excitation light on samples have rarely been investigated. In this paper, by simulating multiple subcellular structure with the distinct refractive index (RI) from water, we study how different thicknesses of this subcellular structure scatter incident light on its optical path of SIM excitation. Because aberrant interference light aggravates with the increase in sample thickness, the reconstruction of the 2D-SIM SR image degraded with the change of focus along the axial axis. Therefore, this work may guide the future development of algorithms to suppress SIM artifacts caused by scattering in thick samples.


Author(s):  
Yanquan Mo ◽  
Fan Feng ◽  
Heng Mao ◽  
Junchao Fan ◽  
Liangyi Chen

Despite its wide application in live-cell super-resolution (SR) imaging, structured illumination microscopy (SIM) suffers from aberrations caused by various sources. Although artefacts generated from inaccurate reconstruction parameter estimation and noise amplification can be minimized, aberrations due to the scattering of excitation light on samples have rarely been investigated. In this paper, by simulating multiple subcellular structure with the distinct refractive index from water, we study how different thicknesses of this subcellular structure scatter incident light on its optical path of SIM excitation. Because aberrant interference light aggravates with the increase in sample thickness, the reconstruction of the 2D-SIM SR image degraded with the change of focus along the axial axis. Therefore, this work may guide the future development of algorithms to suppress SIM artefacts caused by scattering in thick samples. This article is part of the Theo Murphy meeting issue ‘Super-resolution structured illumination microscopy (part 1)'.


Methods ◽  
2015 ◽  
Vol 75 ◽  
pp. 61-68 ◽  
Author(s):  
Laure-Anne Ligeon ◽  
Nicolas Barois ◽  
Elisabeth Werkmeister ◽  
Antonino Bongiovanni ◽  
Frank Lafont

ACS Photonics ◽  
2021 ◽  
Author(s):  
Alice Sandmeyer ◽  
Mario Lachetta ◽  
Hauke Sandmeyer ◽  
Wolfgang Hübner ◽  
Thomas Huser ◽  
...  

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Karin Legerstee ◽  
Tsion E. Abraham ◽  
Wiggert A. van Cappellen ◽  
Alex L. Nigg ◽  
Johan A. Slotman ◽  
...  

AbstractFocal adhesions (FAs) are flat elongated structures that mediate cell migration and link the cytoskeleton to the extracellular matrix. Along the vertical axis FAs were shown to be composed of three layers. We used structured illumination microscopy to examine the longitudinal distribution of four hallmark FA proteins, which we also used as markers for these layers. At the FA ends pointing towards the adherent membrane edge (heads), bottom layer protein paxillin protruded, while at the opposite ends (tails) intermediate layer protein vinculin and top layer proteins zyxin and VASP extended further. At the tail tips, only intermediate layer protein vinculin protruded. Importantly, head and tail compositions were altered during HGF-induced scattering with paxillin heads being shorter and zyxin tails longer. Additionally, FAs at protruding or retracting membrane edges had longer paxillin heads than FAs at static edges. These data suggest that redistribution of FA-proteins with respect to each other along FAs is involved in cell movement.


2021 ◽  
Vol 11 (7) ◽  
pp. 3023
Author(s):  
Kejun Yang ◽  
Chenhaolei Han ◽  
Jinhua Feng ◽  
Yan Tang ◽  
Zhongye Xie ◽  
...  

The surface and thickness distribution measurement for transparent film is of interest for electronics and packaging materials. Structured illumination microscopy (SIM) is a prospective technique for measuring film due to its high accuracy and efficiency. However, when the distance between adjacent layers becomes close, the peaks of the modulation depth response (MDR) start to overlap and interfere with the peak extraction, which restricts SIM development in the field of film measurement. In this paper, an iterative peak separation algorithm is creatively applied in the SIM-based technique, providing a precise peak identification even as the MDR peaks overlap and bend into one. Compared with the traditional method, the proposed method has a lower detection threshold for thickness. The experiments and theoretical analysis are elaborated to demonstrate the feasibility of the mentioned method.


2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Liliana Barbieri ◽  
Huw Colin-York ◽  
Kseniya Korobchevskaya ◽  
Di Li ◽  
Deanna L. Wolfson ◽  
...  

AbstractQuantifying small, rapidly evolving forces generated by cells is a major challenge for the understanding of biomechanics and mechanobiology in health and disease. Traction force microscopy remains one of the most broadly applied force probing technologies but typically restricts itself to slow events over seconds and micron-scale displacements. Here, we improve >2-fold spatially and >10-fold temporally the resolution of planar cellular force probing compared to its related conventional modalities by combining fast two-dimensional total internal reflection fluorescence super-resolution structured illumination microscopy and traction force microscopy. This live-cell 2D TIRF-SIM-TFM methodology offers a combination of spatio-temporal resolution enhancement relevant to forces on the nano- and sub-second scales, opening up new aspects of mechanobiology to analysis.


Sign in / Sign up

Export Citation Format

Share Document