New Method for Multiple Immunodetection on Resin Ultrathin Section in the Field Emission Scanning Electron Microscope

Wandrol, Petr; Vancová, Marie; Nebesářová, Jana

doi:10.1017/s143192761400806x

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“…Next, samples can be imaged in SEM at different accelerating voltage. A higher accelerating voltage enlarge the interaction volume from which the electrons are scattered [ 14 – 16 ]. In ultrathin sections, SEM working in the transmission mode at higher accelerating voltage provide a transmission image of cell structures similarly to TEM [ 16 , 17 ].…”

Section: Introductionmentioning

confidence: 99%

“…A higher accelerating voltage enlarge the interaction volume from which the electrons are scattered [ 14 – 16 ]. In ultrathin sections, SEM working in the transmission mode at higher accelerating voltage provide a transmission image of cell structures similarly to TEM [ 16 , 17 ]. At accelerating voltage below 5 kV, SEM provides detailed information on the surface morphology with minimal charging and radiation damage of sensitive (either dried or hydrated) biological samples.…”

Section: Introductionmentioning

confidence: 99%

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Correlative Fluorescence and Scanning Electron Microscopy of Labelled Core Fucosylated Glycans Using Cryosections Mounted on Carbon-Patterned Glass Slides

Vancová

Nebesářová

2015

PLoS ONE

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The aim of the study is co-localization of N-glycans with fucose attached to N-acetylglucosamine in α1,3 linkage, that belong to immunogenic carbohydrate epitopes in humans, and N-glycans with α1,6-core fucose typical for mammalian type of N-linked glycosylation. Both glycan epitopes were labelled in cryosections of salivary glands isolated from the tick Ixodes ricinus. Salivary glands secrete during feeding many bioactive molecules and influence both successful feeding and transmission of tick-borne pathogens. For accurate and reliable localization of labelled glycans in both fluorescence and scanning electron microscopes, we used carbon imprints of finder or indexed EM grids on glass slides. We discuss if the topographical images can provide information about labelled structures, the working setting of the field-emission scanning electron microscope and the influence of the detector selection (a below-the-lens Autrata improved YAG detector of back-scattered electrons; in-lens and conventional Everhart-Thornley detectors of secondary electrons) on the imaging of gold nanoparticles, quantum dots and osmium-stained membranes.

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