scholarly journals Infrared nanoscopy and tomography of intracellular structures

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
Katerina Kanevche ◽  
David J. Burr ◽  
Dennis J. Nürnberg ◽  
Pascal K. Hass ◽  
Andreas Elsaesser ◽  
...  

AbstractAlthough techniques such as fluorescence-based super-resolution imaging or confocal microscopy simultaneously gather both morphological and chemical data, these techniques often rely on the use of localized and chemically specific markers. To eliminate this flaw, we have developed a method of examining cellular cross sections using the imaging power of scattering-type scanning near-field optical microscopy and Fourier-transform infrared spectroscopy at a spatial resolution far beyond the diffraction limit. Herewith, nanoscale surface and volumetric chemical imaging is performed using the intrinsic contrast generated by the characteristic absorption of mid-infrared radiation by the covalent bonds. We employ infrared nanoscopy to study the subcellular structures of eukaryotic (Chlamydomonas reinhardtii) and prokaryotic (Escherichia coli) species, revealing chemically distinct regions within each cell such as the microtubular structure of the flagellum. Serial 100 nm-thick cellular cross-sections were compiled into a tomogram yielding a three-dimensional infrared image of subcellular structure distribution at 20 nm resolution. The presented methodology is able to image biological samples complementing current fluorescence nanoscopy but at less interference due to the low energy of infrared radiation and the absence of labeling.

2021 ◽  
Author(s):  
Katerina Kanevche ◽  
David Burr ◽  
Andreas Elsaesser ◽  
Pascal-Kolja Hass ◽  
Dennis Nuernberg ◽  
...  

Abstract The few microscopic techniques that simultaneously gather morphological and chemical data often rely on the use of specific markers. To eliminate this flaw, we have developed a method of examining cellular cross sections using the imaging power of scattering-type scanning near-field optical microscopy and Fourier-transform infrared spectroscopy at a spatial resolution far beyond the diffraction limit. Herewith, nanoscale surface and volumetric chemical imaging is performed using the intrinsic contrast generated by the characteristic absorption of mid-infrared radiation by the covalent bonds. We employ infrared nanoscopy to study the subcellular structures of eukaryotic (Chlamydomonas reinhardtii) and prokaryotic (Escherichia coli) species, revealing chemically distinct regions within each cell such as the microtubular structure of the flagellum. Serial 100 nm-thick cellular cross-sections were compiled into a tomogram yielding a three-dimensional infrared image of subcellular structure distribution at 20 nm resolution. The presented methodology is able to image biological samples competing current fluorescence nanoscopy but at less interference due to the low energy of infrared radiation and the absence of labeling.


AIP Advances ◽  
2015 ◽  
Vol 5 (8) ◽  
pp. 084901 ◽  
Author(s):  
Shangting You ◽  
Cuifang Kuang ◽  
Shuai Li ◽  
Xu Liu ◽  
Zhihua Ding

2010 ◽  
Vol 96 (2) ◽  
pp. 023114 ◽  
Author(s):  
B. D. F. Casse ◽  
W. T. Lu ◽  
Y. J. Huang ◽  
E. Gultepe ◽  
L. Menon ◽  
...  

2021 ◽  
Author(s):  
Anna Loeschberger ◽  
Yauheni Novikau ◽  
Ralf Netz ◽  
Marie-Christin Spindler ◽  
Ricardo Benavente ◽  
...  

Three-dimensional (3D) multicolor super-resolution imaging in the 50-100 nm range in fixed and living cells remains challenging. We extend the resolution of structured illumination microscopy (SIM) by an improved nonlinear iterative reconstruction algorithm that enables 3D multicolor imaging with improved spatiotemporal resolution at low illumination intensities. We demonstrate the performance of dual iterative SIM (diSIM) imaging cellular structures in fixed cells including synaptonemal complexes, clathrin coated pits and the actin cytoskeleton with lateral resolutions of 60-100 nm with standard fluorophores. Furthermore, we visualize dendritic spines in 70 micrometer thick brain slices with an axial resolution < 200 nm. Finally, we image dynamics of the endoplasmatic reticulum and microtubules in living cells with up to 255 frames/s.


2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Ralph Götz ◽  
Tobias C. Kunz ◽  
Julian Fink ◽  
Franziska Solger ◽  
Jan Schlegel ◽  
...  

AbstractExpansion microscopy (ExM) enables super-resolution imaging of proteins and nucleic acids on conventional microscopes. However, imaging of details of the organization of lipid bilayers by light microscopy remains challenging. We introduce an unnatural short-chain azide- and amino-modified sphingolipid ceramide, which upon incorporation into membranes can be labeled by click chemistry and linked into hydrogels, followed by 4× to 10× expansion. Confocal and structured illumination microscopy (SIM) enable imaging of sphingolipids and their interactions with proteins in the plasma membrane and membrane of intracellular organelles with a spatial resolution of 10–20 nm. As our functionalized sphingolipids accumulate efficiently in pathogens, we use sphingolipid ExM to investigate bacterial infections of human HeLa229 cells by Neisseria gonorrhoeae, Chlamydia trachomatis and Simkania negevensis with a resolution so far only provided by electron microscopy. In particular, sphingolipid ExM allows us to visualize the inner and outer membrane of intracellular bacteria and determine their distance to 27.6 ± 7.7 nm.


2012 ◽  
Vol 125 (19) ◽  
pp. 4630-4639 ◽  
Author(s):  
M. P. C. van de Corput ◽  
E. de Boer ◽  
T. A. Knoch ◽  
W. A. van Cappellen ◽  
A. Quintanilla ◽  
...  

2012 ◽  
Vol 20 (5) ◽  
pp. 4957 ◽  
Author(s):  
Ignacio Izeddin ◽  
Mohamed El Beheiry ◽  
Jordi Andilla ◽  
Daniel Ciepielewski ◽  
Xavier Darzacq ◽  
...  

1989 ◽  
Vol 206 ◽  
pp. 375-404 ◽  
Author(s):  
Michio Hayakawa ◽  
Fazle Hussain

This paper describes a quantitative study of the three-dimensional nature of organized motions in a turbulent plane wake. Coherent structures are detected from the instantaneous, spatially phase-correlated vorticity field using certain criteria based on size, strength and geometry of vortical structures. With several combinations of X-wire rakes, vorticity distributions in the spanwise and transverse planes are measured in the intermediate region (10d [les ] x [les ] 40d) of the plane turbulent wake of a circular cylinder at a Reynolds number of 13000 based on the cylinder diameter d. Spatial correlations of smoothed vorticity signals as well as phase-aligned ensemble-averaged vorticity maps over structure cross-sections yield a quantitative measure of the spatial coherence and geometry of organized structures in the fully turbulent field. The data demonstrate that the organized structures in the nominally two-dimensional wake exhibit significant three-dimensionality even in the near field. Using instantaneous velocity and vorticity maps as well as correlations of vorticity distributions in different planes, some topological features of the dominant coherent structures in a plane wake are inferred.


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