scholarly journals 3D correlative light and electron microscopy of cultured cells using serial blockface scanning electron microscopy

2016 ◽  
Vol 130 (1) ◽  
pp. 278-291 ◽  
Author(s):  
Matthew R. G. Russell ◽  
Thomas R. Lerner ◽  
Jemima J. Burden ◽  
David O. Nkwe ◽  
Annegret Pelchen-Matthews ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Cultured Cells ◽  
Light And Electron Microscopy ◽  
Scanning Electron

Two new Stauroneis species (Bacillariophyta, Stauroneidaceae) from midwestern karstic Brazilian formations

Phytotaxa ◽  
2018 ◽  
Vol 358 (3) ◽  
pp. 265 ◽  
Author(s):  
EDUARDO A. TUSSET ◽  
PRISCILA I. TREMARIN ◽  
THELMA A.V. LUDWIG
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Light And Electron Microscopy ◽  
Similar Species ◽  
Scanning Electron ◽  
Bow Tie

The morphology of three Stauroneis species from midwestern karstic Brazilian formations was illustrated under light and electron microscopy. Two new Stauroneis are described and compared with similar species in the genus. Stauroneis cavalcantei Tremarin, Tusset & T. Ludwig is characterized by lanceolate valves with subtly protracted apices, and S. karstica Tusset, Tremarin & T. Ludwig by rhombic-lanceolate valves with attenuate to acutely-rounded apices, reduced pseudosepta, bow-tie-shaped stauros, and areolae depressed along prominent longitudinal-oriented apical ribs. Stauroneis alabamae is a scarcely known species and unprecedented scanning electron microscopy illustrations are provided.

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.

Mechanisms Controlling Dormancy in the Arid Zone Grass Aristida contorta. II. Anatomy of the Hull

1974 ◽  
Vol 22 (4) ◽  
pp. 647 ◽  
Author(s):  
JJ Mott ◽  
PW Tynan
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Gas Permeability ◽  
Arid Zone ◽  
Light And Electron Microscopy ◽  
Anatomical Change ◽  
Definitive Statement ◽  
Scanning Electron

The anatomy of the hull directly over the embryo was examined by light and electron microscopy in long-term dormant and non-dormant grains of A. contorta. A lipid-containing layer was noted, covering the surface of the inner epidermis of the hull, and examination by scanning electron microscopy showed that although the layer was intact in all dormant grain examined, it was fractured in non-dormant grain. A definitive statement on the function of these cracks is not possible, but they appear to be an anatomical change leading to increased gas permeability of the hull of non-dormant grains.

scholarly journals X-Ray Microscopy as an Approach to Increasing Accuracy and Efficiency of Serial Block-Face Imaging for Correlated Light and Electron Microscopy of Biological Specimens

2014 ◽  
Vol 21 (1) ◽  
pp. 231-238 ◽  
Author(s):  
Eric A. Bushong ◽  
Donald D. Johnson ◽  
Keun-Young Kim ◽  
Masako Terada ◽  
Megumi Hatori ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Light And Electron Microscopy ◽  
Three Dimensional ◽  
Microscopy Study ◽  
Electron Microscopic ◽  
Intense Staining ◽  
X Ray ◽  
Block Face ◽  
Scanning Electron

AbstractThe recently developed three-dimensional electron microscopic (EM) method of serial block-face scanning electron microscopy (SBEM) has rapidly established itself as a powerful imaging approach. Volume EM imaging with this scanning electron microscopy (SEM) method requires intense staining of biological specimens with heavy metals to allow sufficient back-scatter electron signal and also to render specimens sufficiently conductive to control charging artifacts. These more extreme heavy metal staining protocols render specimens light opaque and make it much more difficult to track and identify regions of interest (ROIs) for the SBEM imaging process than for a typical thin section transmission electron microscopy correlative light and electron microscopy study. We present a strategy employing X-ray microscopy (XRM) both for tracking ROIs and for increasing the efficiency of the workflow used for typical projects undertaken with SBEM. XRM was found to reveal an impressive level of detail in tissue heavily stained for SBEM imaging, allowing for the identification of tissue landmarks that can be subsequently used to guide data collection in the SEM. Furthermore, specific labeling of individual cells using diaminobenzidine is detectable in XRM volumes. We demonstrate that tungsten carbide particles or upconverting nanophosphor particles can be used as fiducial markers to further increase the precision and efficiency of SBEM imaging.

scholarly journals A modified ‘NanoSuit®’ preserves wet samples in high vacuum: direct observations on cells and tissues in field-emission scanning electron microscopy

2017 ◽  
Vol 4 (3) ◽  
pp. 160887 ◽  
Author(s):  
Yasuharu Takaku ◽  
Hiroshi Suzuki ◽  
Hideya Kawasaki ◽  
Isao Ohta ◽  
Daisuke Ishii ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Field Emission ◽  
Structural Changes ◽  
Cultured Cells ◽  
High Vacuum ◽  
Insect Larvae ◽  
Direct Observations ◽  
High Vacuum Environment ◽  
Scanning Electron

Although field-emission scanning electron microscopy (FE-SEM) has proven very useful in biomedical research, the high vacuum required (10 −3 to 10 −7  Pa) precludes direct observations of living cells and tissues at high resolution and often produces unwanted structural changes. We have previously described a method that allows the investigator to keep a variety of insect larvae alive in the high vacuum environment of the electron microscope by encasing the organisms in a thin, vacuum-proof suit, the ‘NanoSuit®'. However, it was impossible to protect wet tissues freshly excised from intact organisms or cultured cells. Here we describe an improved ‘NanoSuit' technique to overcome this limitation. We protected the specimens with a surface shield enhancer (SSE) solution that consists of glycerine and electrolytes and found that the fine structure of the SSE-treated specimens is superior to that of conventionally prepared specimens. The SSE-based NanoSuit affords a much stronger barrier to gas and/or liquid loss than the previous NanoSuit did and, since it allows more detailed images, it could significantly help to elucidate the ‘real' organization of cells and their functions.

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