The structural morphology of the synthesized samples was studied, using Field Emission Scanning Electron Microscope (FESEM), model LEO 435VP from Carl Zeiss™ AG (Jena, Germany). The images were taken at an energy of 10–15 kV. Prior to imaging, the samples were gold-coated, using sputter Q150/ S, Quorum Technologies™ (Darmstadt, Germany). The elemental compositional analysis was performed by EDS (LEO 435VP, Carl Zeiss™ AG (Jena, Germany) at 15 kV.
Leo 435 vp
Manufactured by Zeiss
Sourced in Germany
The LEO 435 VP is a scanning electron microscope (SEM) designed for high-performance imaging and analysis. It features a variable pressure capability, allowing for the examination of non-conductive samples without the need for additional preparation. The core function of the LEO 435 VP is to provide high-resolution imaging and detailed information about the surface topography and composition of a wide range of materials and samples.
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Lab products found in correlation
Med 010, by Oerlikon Balzers (2 mentions)
Sigma 300, by Zeiss (2 mentions)
Aztecenergy system, by Oxford Instruments (1 mentions)
Merlin vp compact, by Zeiss (1 mentions)
Zetasizer nano z, by Malvern Panalytical (1 mentions)
Zetasizer nano s, by Malvern Panalytical (1 mentions)
Cpd 030, by Oerlikon Balzers (1 mentions)
Optiphot, by Nikon (1 mentions)
Jfc 1100, by JEOL (1 mentions)
Helios nanolab 660, by Thermo Fisher Scientific (1 mentions)
Ultra plus, by Zeiss (1 mentions)
Epoxycure, by Buehler (1 mentions)
Scd 050, by Oerlikon Balzers (1 mentions)
Isomet, by Buehler (1 mentions)
20 protocols using leo 435 vp
1
Morphological and Elemental Analysis of Synthesized Samples
2
High-Resolution Scanning Electron Microscopy
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Scanning electron microscopy was carried out with a LEO 435VP at the Geology Department of Trier University and a Carl Zeiss MERLIN VP compact at the IOW, Warnemünde. In both cases, samples were vacuum sputter-coated with gold and an acceleration voltage of 15 kV was applied to produce high-resolution images in back-scattered electron (BSE) mode. At the IOW, EDS analyses (point analyses, area measurements and element mappings) were performed with an Oxford Instruments AztecEnergy system equipped with a X-MAXN80 SDD detector. Elements were detected on the Kα line with a spatial resolution of 1 nm and an energy range of 20 keV. The residence time for each pixel of the BSE images was set to 60 ms for element mappings/area measurements and to 15 ms for single-point analyses, respectively. The EDS measurements were calibrated with various natural and synthetic standards. At concentrations < 1 wt.%, the quantification via EDS yields an increasing relative error (> 70%78 ). Therefore, analyses with concentrations < 1 wt.% were discarded.
3
Characterization of Gelatin Nanospheres
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The unlabeled and labeled gelatin nanospheres were sputter-coated with gold, and the morphology of these gelatin nanospheres was examined using scanning electron microscopy (SEM; LEO 435 VP; Zeiss, Jena, Germany; Sigma-300, Zeiss). The zeta potential of the gelatin nanospheres was characterized by laser Doppler electrophoresis using a Zetasizer Nano-Z instrument (Malvern Instruments Ltd., Worcestershire, UK) in PBS buffer and Milli-Q water. The size of the gelatin nanospheres in swollen state was examined in 154 mM PBS at pH 7.4 and Milli-Q water at pH 7.0 by dynamic light scattering (DLS; Zetasizer Nano-S, Malvern Instruments Ltd., Worcestershire, UK).
4
SEM Analysis of Animal Cell Ultrastructure
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One CP from an animal in each group was excised and immersed for 8 h at 4°C in a 2.5% glutaraldehyde modified Karnovsky solution with 2% paraformaldehyde in 1 M sodium phosphate buffer (pH 7.3). After a rinse with distilled water, the specimens were immersed in a NaOH solution (Ohtani, 1987) for 3 to 5 days at room temperature. The samples were then washed with distilled water for 12 h at 4°C and post-fixed in 1% osmium tetroxide for 2 h at the same temperature. This was followed by a washing with distilled water for 5 h, a dehydration using an ascending alcohol series (70% to absolute) and a drying in a critical point apparatus (Balzers CPD-030) using liquid CO2. The sections were placed on metal stubs, covered with gold (Au) and examined with a Zeiss scanning electron microscope (Leo 435 VP).
5
SEM Analysis of Corn Root Structures
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The roots of inoculated and non-inoculated corn were cut, rinsed in tap water and fixed in 3% glutaraldehyde in 50 mM cacodylate buffer (pH 7.0). Tissue dehydration was performed in a graded series ending in absolute acetone, followed by critical point drying. Then, the samples were mounted on stubs and subjected to gold sputtering (SDC-050 Sputter Coater, BAL-TEC). Finally, the samples were analyzed on a Zeiss LEO 435 VP scanning electron microscope.
6
Somatic Embryogenesis Analysis Protocol
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The samples of embryogenic callus were fixed with FAA, which is a mixture of formalin, glacial acetic acid and 70% ethanol (5:5:90). The tissue was dehydrated in an ethanol series and embedded in paraffin according to the annotation provided by Johansen (1940 ). Using rotatory microtome (Spencer, USA) equipped with a steel knife, fine sections of about 8 µm were made and placed on a glass slide, de-waxed, followed by staining with 5% hematoxylin and 2% eosin dye, and mounted in Canada balsam. Prepared slides were photographed after observing under a light microscope (Nikon Optiphot, Japan).
To know the origin, development and surface morphology of somatic embryos, the SEM was performed. The developing embryos and callus tissues were fixed in 2.0% glutaraldehyde and 2.0% formaldehyde and maintained in pH of 6.8 in 0.1 M of phosphate buffer for 24 h at 4 °C. The tissue was again washed in buffer and later fixed for 2 h in a similar supported 1.0% osmium tetroxide, dried out in an ethanol arrangement and ultimately covered with gold palladium. The tests were performed and captured in an LEO-435VP by Zeiss, Oberkochen, Germany using an electron magnifying lens (operational under 20–25 kV).
To know the origin, development and surface morphology of somatic embryos, the SEM was performed. The developing embryos and callus tissues were fixed in 2.0% glutaraldehyde and 2.0% formaldehyde and maintained in pH of 6.8 in 0.1 M of phosphate buffer for 24 h at 4 °C. The tissue was again washed in buffer and later fixed for 2 h in a similar supported 1.0% osmium tetroxide, dried out in an ethanol arrangement and ultimately covered with gold palladium. The tests were performed and captured in an LEO-435VP by Zeiss, Oberkochen, Germany using an electron magnifying lens (operational under 20–25 kV).
7
Scaffold Preparation for SEM Imaging
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SEM was performed as described earlier57 (link). Briefly, scaffolds were fixed with 3.8% formaldehyde in PBS for at least 1 h and rinsed twice with PBS. Then, the scaffolds were dehydrated by rinsing through graded ethanol/water mixtures (50%, 70%, 80%, 90%, and 100%; each step for 10 min at room temperature). Next, ethanol was slowly exchanged by liquid CO2 (critical point dryer; Balzers CPD, Balter's Union) and the samples were dried using the critical point method. For cross-sections, the specimens were sectioned transversely. Finally, whole tissue and sections were sputtered with a thin layer of gold of ∼10 nm in thickness (sputter coater, Balzers SLD; Balzers Union), and documented by a Leo 435VP (Zeiss) scanning electron microscope.
8
Kidney Ultrastructural Examination
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Kidney samples were fixed in 4% neutral buffered formaldehyde/ 1% glutaraldehyde (pH 7.4) overnight at 4°C, then washed in 0.1 M phosphate buffer, post fixed in 1% Zetterqvist’s osmium tetroxide for 30 minutes, dehydrated with graded ethyl alcohols, and dried in a critical point dryer. Fractured sections were mounted, sputter-coated with gold and viewed with a Zeiss Leo 435 VP scanning electron microscope in the secondary electrons mode for topographical imaging [20 ]. The representative photomicrographs were taken with a digital camera.
9
Shoot Tip Morphological and Anatomical Analysis
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To characterize the morphology, three shoot tips with approximate length of 1 mm, submitted to each of the culture times (30, 45 and 60 days), were fixed in modified Karnovsky's solution (Karnovsky 1965) [glutaraldehyde (2%), paraformaldehyde (2%), CaCl 2 (0.001 M), sodium cacodylate buffer (0.05 M), at pH 7.2] for 48 hours and then dehydrated in an ethyl series (35-100%). The samples were then dried to critical point with liquid CO 2 and mounted on metal supports and sputtered with gold. The images were obtained with a scanning electron microscope with variable pressure (LEO 435 VP, Carl Zeiss, Jena, Germany).
For anatomical characterization, five tips under the same conditions were collected and fixed in the same modified Karnovsky's solution (Karnovsky 1965) for 48 hours, infiltrated and embedded in resin using the Historesin kit (hydroxyethyl methacrylate, Leica, Heidelberg, Germany). The resin was polymerized at room temperature for 48 hours and then serial histological sections (4-5 µm) were obtained with a Leitz model 1516 rotary microtome, placed on slides and stained with acid fuchsin (0,1% p/v), followed by toluidine blue (0.05% p/v) (Feder and O' Brien 1968 ). The slides were observed and photographed with a fluorescence microscope system (B x S1, Olympus Latin America Inc.).
For anatomical characterization, five tips under the same conditions were collected and fixed in the same modified Karnovsky's solution (Karnovsky 1965) for 48 hours, infiltrated and embedded in resin using the Historesin kit (hydroxyethyl methacrylate, Leica, Heidelberg, Germany). The resin was polymerized at room temperature for 48 hours and then serial histological sections (4-5 µm) were obtained with a Leitz model 1516 rotary microtome, placed on slides and stained with acid fuchsin (0,1% p/v), followed by toluidine blue (0.05% p/v) (Feder and O' Brien 1968 ). The slides were observed and photographed with a fluorescence microscope system (B x S1, Olympus Latin America Inc.).
10
Sheep Bone Biopsy Scanning Electron Microscopy
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Biopsies were taken from the iliac crest of adult sheep (∼10 years old; approved by Danish Animal Experiments and Inspectorates, 2011/561-1959) from the study described in Andreasen et al.24 (link) The bone samples were fixed in 4% buffered formaldehyde for 24 h, before transferring to 0.1% buffered formaldehyde. The biopsies were processed according to a previously published procedure25 (link) using a Jeol fine coat gold ion sputter JFC-1100. Scanning electron images were made using a LEO 435VP (Zeiss) operated in a secondary electron mode with a 300 V positive bias at the detector. The working distance was 35 mm to assure a large depth of field in the images.
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