Cells were bathed in a solution containing 2% glutaraldehyde, 2% paraformal-dehyde and 0.05 M sodium cacodylate (pH 7.2) (all from Electron Microscopy Sciences) for 2 h at 4˚C. The cells were then fixed by incubation with 1% osmium tetroxide (Electron Microscopy Sciences) for 1 h at 4˚C, dehydrated with increasing concentrations of ethanol (25, 50, 70, 90 and 100%) for 5 min at each concentration, and embedded in epoxy resin (Embed 812; Electron Microscopy Sciences) for 48 h at 60˚C following the manufacturers' instructions. Ultrathin sections (60-nm-thick) were sliced using an LKB-III ultratome (Leica Microsystems GmbH). The slices were stained with 0.5% uranyl acetate (Electron Microscopy Sciences) for 20 min and with 0.1% lead citrate (Electron Microscopy Sciences) for 7 min at room temperature. Fluorescent images were recorded using the Hitachi H7650 electron microscope (Hitachi, Ltd.; magnification, ×10,000) located at the Center for University-Wide Research Facilities (CURF) at Jeonbuk National University.
Lkb 3 ultratome
Manufactured by Leica Microsystems
Sourced in Germany
The LKB-III ultratome is a precision instrument designed for the preparation of ultrathin sections for electron microscopy. It utilizes a diamond or glass knife to cut samples into extremely thin slices, typically ranging from 50 to 100 nanometers in thickness. The LKB-III is a reliable and versatile tool for researchers and laboratories requiring high-quality sample preparation for advanced microscopic analysis.
Automatically generated - may contain errors
Lab products found in correlation
Osmium tetroxide, by Electron Microscopy Sciences (8 mentions)
Embed 812, by Electron Microscopy Sciences (7 mentions)
Glutaraldehyde, by Electron Microscopy Sciences (7 mentions)
H 7650 electron microscope, by Hitachi (7 mentions)
Lead citrate, by Electron Microscopy Sciences (6 mentions)
Uranyl acetate, by Electron Microscopy Sciences (6 mentions)
0.05 m sodium cacodylate, by Electron Microscopy Sciences (3 mentions)
Paraformaldehyde (pfa), by Electron Microscopy Sciences (2 mentions)
M sodium cacodylate, by Electron Microscopy Sciences (2 mentions)
Uranyl acetate, by Ted Pella (1 mentions)
Lead citrate, by Ted Pella (1 mentions)
H 7600, by Hitachi (1 mentions)
Sodium cacodylate, by Electron Microscopy Sciences (1 mentions)
Embed812 epoxy resin, by Electron Microscopy Sciences (1 mentions)
10 protocols using lkb 3 ultratome
1
Electron Microscopy Specimen Preparation
2
Huh7 Cell Ultrastructure Analysis
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Following fixation of Huh7 cells in 2% glutaraldehyde (Electron Microscopy Sciences, Hatfield, PA, USA) and 2% paraformaldehyde (EMS, USA) in 0.05 M sodium cacodylate (pH 7.2; Electron Microscopy Sciences) for 2 h at 4°C, specimens were fixed in 1% osmium tetroxide (Electron Microscopy Sciences) for 1 h at 4°C, dehydrated with increasing ethanol (25, 50, 70, 90 and 100%) for 5 min each at 4°C and embedded in epoxy resin (Embed 812; Electron Microscopy Sciences) for 48 h at 60°C according to the manufacturers' instructions. Ultrathin sections (60 nm) were prepared using an LKB-III ultratome (Leica Microsystems GmbH, Wetzlar, Germany) and were stained with 0.5% uranyl acetate (Electron Microscopy Sciences) for 20 min and 0.1% lead citrate (Electron Microscopy Sciences) for 7 min at room temperature. Images were recorded on a Hitachi H7650 electron microscope (magnification, ×10,000; Hitachi, Ltd., Tokyo, Japan) installed at the Center for University-Wide Research Facilities (CURF) at Chonbuk National University.
3
Electron Microscopy Sample Preparation
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The specimens were fixed in 2.5% glutaraldehyde and post-fixed in 1% osmium tetroxide, dehydrated through a graded ethanol series, and embedded in epoxy resin. Serial ultrathin sections were cut to 70 µm on an LKB-III ultratome (Leica Microsystems GmbH, Wetzlar, Germany), stained with uranyl acetate (Ted Pella, Inc., Redding, CA, USA) and lead citrate (Ted Pella, Inc.), and examined on an electron microscope (H7600; Hitachi, Tokyo, Japan) at an acceleration voltage of 100 kV.
4
Ultrastructural Analysis of Trypsinized Cells
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Trypsinized cells were fixed with 2% glutaraldehyde (Electron Microscopy Sciences) for 2 h at 4°C in PBS, followed by 2% osmium tetroxide (Electron Microscopy Sciences), and dehydrated with an ethanol series (25, 50, 70, 90 and 100%) for 5 min each. After dehydration, the samples were embedded in epoxy resin (Embed 812; Electron Microscopy Sciences) for 48 h at 60°C according to the manufacturer's instructions. Ultrathin sections (60 nm) were prepared using an LKB III ultratome (Leica Microsystems GmbH) and stained with 0.5% uranyl acetate (Electron Microscopy Sciences) for 20 min and 0.1% lead citrate (Electron Microscopy Sciences) for 7 min at RT. Images were captured on a Hitachi H7650 electron microscope (magnification ×10,000; Hitachi, Ltd.) installed at the Center for University-Wide Research Facilities (CURF) at Jeonbuk National University (JBNU).
5
Transmission Electron Microscopy of SK-N-SH Cells
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TEM was performed as described previously (Moon and Park, 2020 (link)). Briefly, Trypsinized SK-N-SH cells were fixed with 2% paraformaldehyde and 2% glutaraldehyde in 0.05 M sodium cacodylate, pH 7.2. The cells were post-fixed with 1% osmium tetroxide (Electron Microscopy Sciences). The specimens were dehydrated using an ethanol series (25%, 50%, 70%, 90%, and 100%) for 5 min each. After dehydration, samples were embedded in epoxy resin (Embed 812; Electron Microscopy Sciences) for 48 h at 60 °C as per the manufacturer's instructions. Ultrathin sections (60 nm) were prepared using an LKB III ultratome (Leica Microsystems GmbH) and stained with 0.5% uranyl acetate (Electron Microscopy Sciences) for 20 min and 0.1% lead citrate (Electron Microscopy Sciences) for 7 min at room temperature. Images were recorded on a Hitachi H7650 electron microscope (Hitachi, Ltd., Tokyo, Japan; magnification, x10,000) installed at the Center for University-Wide Research Facilities (CURF) at Jeonbuk National University.
6
Transmission Electron Microscopy Preparation
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Cells were fixed in 2% glutaraldehyde (Electron Microscopy Sciences, Hatfield, PA, USA) and 2% paraformaldehyde (EMS, USA) in 0.05 M sodium cacodylate (pH 7.2; Electron Microscopy Sciences) for 2 h at 4 °C. After, the samples were fixed in 1% osmium tetroxide (Electron Microscopy Sciences) for 1 h at 4 °C, dehydrated by incubating in alcohol solutions of increasing concentration (25, 50, 70, 90 and 100%) for 5 min each and entrenched in epoxy resin (Embed 812; Electron Microscopy Sciences) for 48 h at 60 °C following the manufacturers’ instructions. Ultrathin sections (60 nm) were cut using the LKB-III ultratome (Leica Microsystems GmbH, Wetzlar, Germany) and stained using 0.5% uranyl acetate (Electron Microscopy Sciences) for 20 min and 0.1% lead citrate (Electron Microscopy Sciences) for 7 min at room temperature. Images were captured on a Hitachi H7650 electron microscope (Hitachi, Ltd., Tokyo, Japan; magnification, 10,000×) at the Center for University-Wide Research Facilities (CURF), Chonbuk National University.
7
Transmission Electron Microscopy Sample Preparation
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The cells for TEM were fixed in 2% glutaraldehyde [Electron Microscopy Sciences (EMS)] and 2% paraformaldehyde (EMS) in 0.05 M sodium cacodylate (pH 7.2; EMS) for 2 h at 4°C. The samples were fixed in 1% osmium tetroxide (EMS) for 1 h at 4°C, dehydrated with a graded ethanol series (25, 50, 70, 90 and 100%) for 5 min each, and embedded in epoxy resin (Embed 812; EMS) for 48 h at 60°C following to the manufacturer's instructions. Ultrathin sections (60 nm) were cut using an LKB-III Ultratome (Leica Microsystems GmbH) and stained with 0.5% uranyl acetate (EMS) for 20 min and 0.1% lead citrate (EMS) for 7 min at RT. The fluorescent images were recorded on a Hitachi H7650 electron microscope (Hitachi, Ltd.; magnification, ×10,000) installed at the Center for University-Wide Research Facilities (CURF) at Jeonbuk National University.
8
Ultrastructural Imaging of Cells
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Following the fixation of cells in 2% glutaraldehyde (Electron Microscopy Sciences) and 2% paraformaldehyde (Electron Microscopy Sciences) in 0.05 M sodium cacodylate (pH 7.2; Electron Microscopy Sciences) for 2 h at 4°C, the specimens were fixed in 1% osmium tetroxide (Electron Microscopy Sciences) for 1 h at 4°C, dehydrated in increasing ethanol concentration (25, 50, 70, 90 and 100%) for 5 min each, and embedded in Embed 812 epoxy resin (Electron Microscopy Sciences) for 48 h at 60°C according to the manufacturers' protocol. Ultrathin sections (60 nm) were prepared using a LKBIII ultratome (Leica Microsystems GmbH) and stained with 0.5% uranyl acetate (Electron Microscopy Sciences) for 20 min and 0.1% lead citrate (Electron Microscopy Sciences) for 7 min at room temperature. Images were captured using a Hitachi H7650 electron microscope (Hitachi, Ltd.) at magnification, ×10,000, installed at the Center for University-Wide Research Facilities (CURF), Jeonbuk National University.
9
Transmission Electron Microscopy Sample Preparation
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Cells were xed in 2% glutaraldehyde (Electron Microscopy Sciences, Hat eld, PA, USA) and 2% paraformaldehyde (EMS, USA) in 0.05 M sodium cacodylate (pH 7.2; Electron Microscopy Sciences) for 2 h at 4 °C. After, the samples were xed in 1% osmium tetroxide (Electron Microscopy Sciences) for 1 h at 4 °C, dehydrated by incubating in alcohol solutions of increasing concentration (25, 50, 70, 90 and 100%) for 5 min each and entrenched in epoxy resin (Embed 812; Electron Microscopy Sciences) for 48 h at 60 °C following the manufacturers' instructions. Ultrathin sections (60 nm) were cut using the LKB-III ultratome (Leica Microsystems GmbH, Wetzlar, Germany) and stained using 0.5% uranyl acetate (Electron Microscopy Sciences) for 20 min and 0.1% lead citrate (Electron Microscopy Sciences) for 7 min at room temperature. Images were captured on a Hitachi H7650 electron microscope (Hitachi, Ltd., Tokyo, Japan; magni cation, 10,000×) at the Center for University-Wide Research Facilities (CURF), Chonbuk National University.
10
Transmission Electron Microscopy Sample Preparation
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Cells were xed in 2% glutaraldehyde (Electron Microscopy Sciences, Hat eld, PA, USA) and 2% paraformaldehyde (EMS, USA) in 0.05 M sodium cacodylate (pH 7.2; Electron Microscopy Sciences) for 2 h at 4 °C. After, the samples were xed in 1% osmium tetroxide (Electron Microscopy Sciences) for 1 h at 4 °C, dehydrated by incubating in alcohol solutions of increasing concentration (25, 50, 70, 90 and 100%) for 5 min each and entrenched in epoxy resin (Embed 812; Electron Microscopy Sciences) for 48 h at 60 °C following the manufacturers' instructions. Ultrathin sections (60 nm) were cut using the LKB-III ultratome (Leica Microsystems GmbH, Wetzlar, Germany) and stained using 0.5% uranyl acetate (Electron Microscopy Sciences) for 20 min and 0.1% lead citrate (Electron Microscopy Sciences) for 7 min at room temperature. Images were captured on a Hitachi H7650 electron microscope (Hitachi, Ltd., Tokyo, Japan; magni cation, 10,000×) at the Center for University-Wide Research Facilities (CURF), Chonbuk National University.
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