1×105 RK13 cells were infected with 1×106 spores on glass coverslips imprinted with three location markers. Cells were fixed with 4% paraformaldehyde in PBS for 30 min after three days post-infection. After being washed three times by TBST (TBS plus 0.05%Tween 20), coverslips were blocked by 3% BSA in TBST for 1hr at room temperature. EhPTP4 monoclonal antibody (MAb-EhPTP4; clone F4-6) at a 1:10 dilution and EhPTP1 rabbit polyclonal antibody (rab-PcAb-EhPTP1) at a 1:500 dilution were incubated with cells for 1hr at room temperature. After being washed three times by TBST, Alexa Fluor 488-labeled anti-mouse IgG antibody and Alexa Fluor 594-labeled anti-rabbit IgG antibody were added at 1:500 dilutions. Mouse and rabbit preimmunization sera were used as negative controls. Samples were washed three times by TBST and imaged using a Zeiss AxioObserver microscope equipped with Axiovision software with "shuttle & find" to mark cell locations. After fluorescence imaging, samples were fixed with 2.5% glutaraldehyde, dehydrated in ethanol, critical point dried (Tousimis Samdri 790), and coated with chromium (EMS 150T-ES). The same cells were automatically located in the Zeiss Supra 40 Field Emission Scanning Electron Microscope and imaged with a secondary electron detector. Fluorescence and SEM images were correlated with Zeiss AxioVision.
Supra 40 field emission scanning electron microscope
Manufactured by Zeiss
Sourced in Germany
The Supra 40 Field Emission Scanning Electron Microscope is a high-resolution imaging tool that uses a focused beam of electrons to produce detailed images of small-scale structures. It is designed to capture precise, high-quality images of a wide range of samples.
Automatically generated - may contain errors
6 protocols using supra 40 field emission scanning electron microscope
1
Correlative Fluorescence and SEM Microscopy
2
Scanning Electron Microscopy of SINV-Infected Cells
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Vero cells were cultured on glass coverslips. Cells were mock infected or infected with WT SINV at an MOI of 5 and fixed after 9 h at 37°C. Alternatively, cells were mock transfected or transfected with the plasmids WT+Cp or Y399R+Cp and fixed after 24 h at 37°C. Fixation was for 20 min at room temperature with 4% paraformaldehyde, 2% glutaraldehyde in phosphate-buffered saline. Cells were then fixed with 2.5% glutaraldehyde, postfixed in 2% osmium tetroxide, dehydrated in ethanol, critical-point dried (Tousimis Samdri 790 critical point dryer), and coated with chromium (EMS 150T-ES sputtercoater). Cells were imaged using a Zeiss Supra 40 field emission scanning electron microscope (SEM) using a secondary electron detector.
3
Electrical Characterization of Biosensor Devices
4
Quantitative DNA Detection using GNPs
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The microchips were incubated in a solution of the streptavidin–GNPs conjugate in PBST containing 1% BSA at 37 °C for 30 min, washed twice with PBST for 10 min, once with H2O for 3 min, and dried on air. The GNPs on the silicon surface were detected using a Supra-40 field emission scanning electron microscope (Carl Zeiss AG, Jena, Germany) with an InLens secondary electron detector built in the microscope column. The accelerating voltage and beam current were adopted to achieve the best resolution and contrast for GNP visualization. The number of GNPs on microchip fragments was counted using the Gwyddion software (Czech Metrology Institute, Brno, Czech Republic). The limit of DNA detection (LOD) was calculated as the mean number of GNPs registered for a blank DNA sample (0 pM DNA) plus 2 standard deviations (SD, n = 10).
5
Evaluating Composite Metal-Polymer Foams
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The morphology of the composite metal–polymer system and the foam adhesion to the metal cage were inspected using a Supra 40 Field-Emission Scanning Electron Microscope (Zeiss, Germany). Images were acquired in secondary electron mode at 5.00 kV for macroscopical evaluation of the different composite manufacturing techniques, pure foaming, and electrowetting, while for the analysis of the foamed single strut and its adhesion, a 2.00 kV was imposed. Before the SEM inspection, specimens underwent the sputtering of a platinum-palladium (Pt/Pd, 80:20) conductive thin coating (Q150T ES, Quorum Technologies, Lewes, UK).
6
Correlative Ultrastructural Imaging
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Samples were immersion fixed in 2.0% paraformaldehyde, 2.5% glutaraldehyde in 0.1M sodium cacodylate buffer, then processed using a modified National Center for Microscopy and Imaging method of OTO40 (link). In brief, samples were post fixed with reduced osmium, treated with thiocarbohydrazide, further stained with osmium, en bloc stained with uranyl acetate, further stained with lead aspartate, dehydrated in a graded series of ethanol, and embedded into LX112 resin. 55nm-thick sections were cut on a Leica Artos microtome using a Diatome AT 35° knife, and picked up on freshly glowed silicon wafers. Sections were observed on Zeiss Supra 40 Field Emission Scanning Electron Microscope in backscatter mode, using an acceleration voltage of 8.0 kV.
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