TEM was used to visualize the intracellular distribution of MNP-Mn(II). For this, MNP-Mn(II)-labeled and unlabeled cells were resuspended and centrifuged at 1,500×g for 5 min, yielding cell pellets. After the supernatants were removed, cell pellets were fixed at 4°C with 2% glutaraldehyde for 1 h, and washed with PBS several times. The cells were then post-fixed with 1% OsO4 for 1 h, dehydrated in a graded series of ethanol solutions. The cells in ultrathin sections were obtained using an ultramicrotome (Leica EM UC-6; Leica Microsystems, Mannheim, Germany) and stained with uranyl acetate and lead citrate. Finally, the sections were visualized using a TEM (JEM-1011; JEOL).
Leica em uc6
Manufactured by Leica Microsystems
Sourced in Germany
The Leica EM UC6 is an ultra-microtome designed for ultrathin sectioning of biological and material samples. It features precise thickness control and a motorized system for accurate cutting. The EM UC6 is suitable for a range of applications requiring high-quality ultrathin sections.
Automatically generated - may contain errors
Lab products found in correlation
Jem 1230, by JEOL (3 mentions)
Diamond knife, by Electron Microscopy Sciences (2 mentions)
Embed 812 kit, by Electron Microscopy Sciences (2 mentions)
Jem 1011, by JEOL (1 mentions)
Bx53 microscope, by Olympus (1 mentions)
1011 electron microscope, by JEOL (1 mentions)
Epon 812, by Electron Microscopy Sciences (1 mentions)
Jem 1400, by JEOL (1 mentions)
Dfc 290 hd camera, by Leica camera (1 mentions)
Dmi4000b, by Leica camera (1 mentions)
Esem xl 30 feg, by Philips (1 mentions)
Jem 2100plus tem, by JEOL (1 mentions)
Lr white acrylic resin, by Agar Scientific (1 mentions)
Gatan rio16 camera, by Ametek (1 mentions)
Mira3 feg sem, by TESCAN (1 mentions)
H 7650 tem, by Hitachi (1 mentions)
Jem 1400 plus electron microscope, by JEOL (1 mentions)
Dfc300fx, by Leica camera (1 mentions)
Dm4000b, by Leica camera (1 mentions)
Technovit 7100 resin, by Kulzer (1 mentions)
37 protocols using leica em uc6
1
Visualizing Intracellular Distribution of MNP-Mn(II)
2
TEM Analysis of Autophagy in RF-EMF Exposure
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the TEM studies, the cells were exposed to RF-EMFs (1, 2, or 4 W/kg) for 24 h, harvested, washed twice with PBS (pH 7.4) at room temperature, and fixed with 2.5% glutaraldehyde. The cells were then washed three times with PBS, fixed with 1% osmic acid for 2–3 h, washed three times with PBS, dehydrated, embedded in paraffin, cut into 70-nm-thick sections using an Ultrathin slicing machine (Leica EM UC6; Leica Microsystems, Wetzlar and Mannheim, Germany), and stained with uranyl acetate-lead citrate. The cells were then observed using a transmission electron microscope (JEM1230, JEOL Ltd., Tokyo, Japan) for the detection of autophagic vacuoles.
3
Anther Preparation for Microscopy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Fresh anthers from both wild type and the mutant at different stages were immersed in FAA solution (50% ethanol, 5% glacial acetic acid, 5% formalin) for 24 h at room temperature for fixation for scanning electron microscope (SEM) analysis. The samples were then dehydrated in a serial of ethanol gradients (50–100%). After critical-point drying, the anthers were coated with palladium gold and then observed using a scanning electron microscope (HITACHI S-3400N, Tokyo, Japan).
For cytological observation, the anthers were pricked and fixed in FAA solution overnight. The samples were dehydrated using a serial of ethanol (50–100%) and embedded in spurr resin. Semi-thin sections were obtained while using a Leica UE (Leica Microsystems, Berlin, Germany), stained with 0.05% toluidine blue and observed with an Olympus BX-53 microscope (Tokyo, Japan). For laser scanning confocal microscopy analysis, fresh anthers were vacuum infiltrated and prefixed in 3.5% glutaraldehyde (0.1 M phosphate buffer, pH 7.4), followed by rinsing with 0.1 M phosphate buffer. The samples were transferred into 1% osmium tetraoxide and rinsed with 0.1 M phosphate buffer. After fixation, the samples were dehydrated using an ethanol series from 50% to 100% and then embedded in spurr resin. Ultra-thin sections were collected with a Leica EM-UC6 (Leica Microsystems, Berlin, Germany).
For cytological observation, the anthers were pricked and fixed in FAA solution overnight. The samples were dehydrated using a serial of ethanol (50–100%) and embedded in spurr resin. Semi-thin sections were obtained while using a Leica UE (Leica Microsystems, Berlin, Germany), stained with 0.05% toluidine blue and observed with an Olympus BX-53 microscope (Tokyo, Japan). For laser scanning confocal microscopy analysis, fresh anthers were vacuum infiltrated and prefixed in 3.5% glutaraldehyde (0.1 M phosphate buffer, pH 7.4), followed by rinsing with 0.1 M phosphate buffer. The samples were transferred into 1% osmium tetraoxide and rinsed with 0.1 M phosphate buffer. After fixation, the samples were dehydrated using an ethanol series from 50% to 100% and then embedded in spurr resin. Ultra-thin sections were collected with a Leica EM-UC6 (Leica Microsystems, Berlin, Germany).
4
Ultrastructural Imaging of APEX-Tagged Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Ascorbate peroxidase (APEX) processing of baby hamster kidney (BHK; CCL-10, ATCC) cells was co-transfected with emGFP-Slc37a2 and APEX-GBP (GFP-binding peptide) was performed according to the methods described in ref. 70 (link). Briefly, transfected cells were fixed in 2.5% glutaraldehyde, and washed repeatedly in 0.1 M sodium cacodylate buffer prior to the DAB reaction. Cells were incubated with DAB in the presence of H2O2 for 30 min at RT, washed in 0.1 M sodium cacodylate buffer, post-fixed in 1% osmium tetroxide for 2 min, washed again, then serially dehydrated in increasing percentages of ethanol. Cells were serially infiltrated with LX112 resin in a Pelco Biowave microwave and then polymerized overnight at 60 °C. Ultrathin sections were cut on an ultramicrotome (Leica EM UC6, Leica Microsystems) and imaged using a JEOL1011 electron microscope (JEOL) at 80 kV.
5
Transmission Electron Microscopy Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Transmission EM was performed as described previously13 (link). In brief, cells were fixed with 2.5% glutaraldehyde (GA) in 50 mM sodium cacodylate buffer (CaCo), supplemented with 2% sucrose, 50 mM KCl, 2.6 mM MgCl2 and 2.6 mM CaCl2, for 30 min at room temperature. After five washes with 50 mM CaCo, samples were incubated with 2% OsO4 in 25 mM CaCo for 40 min on ice, washed three times with EM-grade water and incubated in 0.5% uranyl acetate in water overnight at 4 °C. Samples were rinsed three times with EM-grade water, dehydrated in a graded ethanol series (from 40 to 100%) at room temperature, embedded in Epon 812 (Electron Microscopy Sciences) and polymerized for 2 days at 60 °C. After polymerization, ultrathin sections of 70 nm were obtained by sectioning with an ultramicrotome Leica EM UC6 (Leica Microsystems) and mounted on a slot grid. Sections were counterstained using 3% uranyl acetate in 70% methanol for 5 min and lead citrate (Reynold’s) for 2 min and examined by using a JEOL JEM-1400 (JEOL) operating at 80 kV and equipped with a 4 K TemCam F416 (Tietz Video and Image Processing Systems GmbH).
6
Cellular Distribution of Particles by SEM-EDX
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The particle distribution within the cells was investigated after 24 h of exposure to the highest concentration of 50 mg/l. Particle-treated cells were observed by an inverse microscope (Leica DMI 4000B, magnification 200×) connected to a Leica DFC 290 HD camera. Detailed analyses of the localization of particles inside the cells were performed by scanning electron microscopy (SEM, Philips ESEM XL 30 FEG) coupled with energy-dispersive X-ray spectroscopy (EDX with EDAX detecting unit, EDAX Inc.) for element analysis. Therefore, cell pellets containing ~10 × 105 cells were fixated with 2.5 % glutaraldehyde in PBS after exposure. After washing with PBS, the samples were stained with osmium tetroxide, washed with deionized water, dehydrated with a graduated acetone series, including an additional staining step with uranyl acetate, and finally embedded in epoxy resin. In the last step, samples were cut by a microtome (Leica EM UC6, Leica Microsystems GmbH) and measured by SEM-coupled EDX to determine the element distribution (Bastian et al. 2009 (link); Kühnel et al. 2012 (link)).
7
Epicarp Microstructure Characterization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The untreated fruit epicarps were broken into small pieces. Untreated fragments or polished fragments of epicarps were embedded in catalyzed London resin (LR) white acrylic resin (Agar Scientific Ltd.) and hardened overnight at 60 °C. After resin embedding, the embedded sample block was transversely cut with a 45° diamond knife (Diatom) equipped on an ultramicrotome Leica EM UC6 (Leica Microsystems). The sample block was transferred onto a SEM stub and coated with 10 nm Pt for imaging. The SEM (Tescan MIRA3 FEG-SEM) used to observe the top view was operated at 3 kV with a working distance of 5.2 mm and using an in-beam secondary electron detector.
Ultrathin sections with a thickness of 100 nm were produced and transferred onto carbon coated Cu grids (Agar Scientific Ltd). The TEM (Hitachi H-7650 TEM) equipped with an AMT 2k × 2k digital camera system was operated at 80 kV. For the width measurement, ultrathin sections of thickness of 30 to 50 nm were prepared and transferred onto carbon-coated Cu grids. The observation was done in a low-dose condition using a JEM 2100Plus TEM (Jeol Ltd.) operated at 200 kV and equipped with a Gatan Rio16 camera (Gatan, Inc.). ForSI Appendix, Fig. S1 , untreated epicarp fragments were mounted on carbon tape with cross-sections exposed and sputter-coated with 10 nm Au/Pd.
Ultrathin sections with a thickness of 100 nm were produced and transferred onto carbon coated Cu grids (Agar Scientific Ltd). The TEM (Hitachi H-7650 TEM) equipped with an AMT 2k × 2k digital camera system was operated at 80 kV. For the width measurement, ultrathin sections of thickness of 30 to 50 nm were prepared and transferred onto carbon-coated Cu grids. The observation was done in a low-dose condition using a JEM 2100Plus TEM (Jeol Ltd.) operated at 200 kV and equipped with a Gatan Rio16 camera (Gatan, Inc.). For
8
Electron Microscopy of HeLa Cell Transfection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
HeLa cells were grown on ACLAR disks and transfected with either NL4-3 or NL4-3(NC–Fluo-SP2) vectors. Cells were fixed in 2.5% glutaraldehyde plus 1% paraformaldehyde in 0.1 M cacodylic buffer for 30 min and then embedded in resin using an Embed 812 kit (Electron Microscopy Sciences, Hatfield, PA, USA) and sectioned at 80 nm with a diamond knife (Diatome) using a Leica EM UC6 (Leica Microsystems, Wetzlar, Germany). Sections were visualized using a JEM 1400 Plus electron microscope (JEOL, Tokyo, Japan) at 120 kV.
9
Cuticle Thickness Measurement in Olive Pericarp
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Pericarp fruit samples were chopped to little cubes (roughly 2 mm per side) and fixed in a formaldehyde-acetic acid (FAA) solution [5% (v/v) formaldehyde and 5% (v/v) glacial acetic acid in 1:1 (v/v) ethanol-distilled water] for 12 h. Samples were dehydrated in aqueous solutions containing increasing ethanol concentrations up to 100% (v/v). Dehydrated samples were transferred to Eppendorf tubes for infiltration and polymerization in Technovit 7100® resin (Heraeus Kulzer GmbH, Wehrheim, Germany), and the resin was dried at 45°C for 24 h.
Resin-embedded samples were cut in 4-µm-thick sections using an ultramicrotome (Leica EM UC6, Leica Microsystems GmbH, Wetzlar, Germany), and subsequently stained on a slide for 15 min in a Sudan IV lysochrome solution [5% (w/v) in 85% (v/v) ethanol] in order to visualize the lipidic constituents of fruit cuticles. Excess staining was removed by rinsing in 50% (v/v) ethanol, and samples allowed to dry at room temperature. Olive pericarp sections were observed and photographed using a microscope (Leica DM4000 B) with a coupled camera (Leica DFC300 FX). Cuticle thickness was determined from five images obtained from five different fruit per cultivar and maturity stage with the Fiji image processing software (Schindelin et al., 2012 (link)).
Resin-embedded samples were cut in 4-µm-thick sections using an ultramicrotome (Leica EM UC6, Leica Microsystems GmbH, Wetzlar, Germany), and subsequently stained on a slide for 15 min in a Sudan IV lysochrome solution [5% (w/v) in 85% (v/v) ethanol] in order to visualize the lipidic constituents of fruit cuticles. Excess staining was removed by rinsing in 50% (v/v) ethanol, and samples allowed to dry at room temperature. Olive pericarp sections were observed and photographed using a microscope (Leica DM4000 B) with a coupled camera (Leica DFC300 FX). Cuticle thickness was determined from five images obtained from five different fruit per cultivar and maturity stage with the Fiji image processing software (Schindelin et al., 2012 (link)).
10
Preparation of Phloem Tissue Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Thin transversal-sections were cut from phloem cubes embedded in LR White resin (TAAB Laboratories, Aldermason, Berkshire, UK) for routine observations of morphology and presence of fungal hyphae. Samples were processed for light microscopy according to Nagy et al. (2000) (link). Briefly, pieces were fixated (in 2% paraformaldehyde and 1.25% glutaraldehyde in 50 mmol/L L-piperazine-N-N′-bis (2-ethanesulfonic) acid buffer (pH 7.2) for 12 h at room temperature), and dehydrated in an ethanol series (70-80-90-96-4 × 100%) before infiltration and polymerization (at 60 °C for 24 h) with L. R. White acrylic resin. Cross-sections (1 µm thick) were cut from all control samples and all inoculated samples collected 3–35 days after inoculation using an ultramicrotome for resin sections (Leica EM UC6; Leica Microsystems, Wetzlar, Germany). Both resin sections and the cryo-sections collected for morphological examination by light microscopy were dried onto superfrost® Plus glass slides (Menzel-Gläzer®; Thermo Scientific, Gerhard Menzel GmbH, Braunschweig, Germany) and stained with Stevenel’s blue (del Cerro, Cogen & del Cerro, 1980 (link)).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!