The excised livers, cut into small pieces, were immediately fixed by direct immersion in 4% glutaraldehyde (Electron Microscopy Sciences, Hatfield, PA, USA) in phosphate-buffered saline (PBS 0.1 M, pH 7.2) for 48 h at 4 °C. After post-fixation for 2 h with osmium tetroxide (1% in the same buffer) and dehydration in graded ethanol, specimens for light microscopy (LM) and transmission electron microscopy (TEM) were embedded in epoxy resin (Araldite 502/Embed 812, Electron Microscopy Sciences). Semi-thin sections (1 μm) were stained with toluidine blue; these sections were observed and photographed by a LM LEITZ Dialux EB 20 (Leica Microsystems, Wetzlar, Germany). Ultrathin sections (800 Å) were stained with uranyl acetate replacement and contrasted using lead citrate (Electron Microscopy Sciences), then observed by a JEOL 1011 electron microscope (JEOL, Inc., Peabody, MA, USA).
Lm leitz dialux eb 20
Manufactured by Leica
Sourced in Germany
The LM LEITZ Dialux EB 20 is a microscope designed for laboratory use. It features a binocular observation head and a built-in illumination system.
Automatically generated - may contain errors
Lab products found in correlation
Araldite 502 embed 812, by Electron Microscopy Sciences (4 mentions)
Glutaraldehyde, by Electron Microscopy Sciences (3 mentions)
Lead citrate, by Electron Microscopy Sciences (2 mentions)
Ultracut uct, by Leica (2 mentions)
Em10 electron microscope, by Zeiss (2 mentions)
1011 electron microscope, by JEOL (1 mentions)
Epon araldite, by Electron Microscopy Sciences (1 mentions)
4 protocols using lm leitz dialux eb 20
1
Ultrastructural Analysis of Excised Liver
2
Fixation and Embedding of Liver Tissue
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Liver were quickly excised cut into small pieces, and fixed by direct immersion in 4% glutaraldehyde in phosphate-buffered saline (PBS 0.1 M, pH 7.2) for 48 h at 4 °C. After post-fixation for 2 h with osmium tetroxide (1% in the same buffer), samples were dehydration in an increasing series of ethanol and then embedded in epoxy resin (Araldite 502/Embed 812, Electron Microscopy Sciences, Hatfield, PA, USA). Semi-thin sections (1 µm) were stained with toluidine blue, observed and photographed by a LM LEITZ Dialux EB 20 (Leica Microsystems, Wetzlar, Germany) equipped with a digital camera.
3
Electron Microscopy Sample Preparation Protocol
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For conventional light microscopy (LM) and transmission electron microscopy (TEM), tissues were subjected to primary aldehyde fixation in 3% glutaraldehyde (in 0.1M phosphate buffer, pH 7.2; (Electron Microscopy Sciences, Hatfield, PA, USA) for 3 h at 4°C followed by postfixation in in buffered 1% osmium tetroxide. The samples were then dehydrated through a graded series of ethanol, soaked in propylene oxide and embedded in Epon‐Araldite (Araldite 502/Embed 812, Electron Microscopy Sciences).
Blocks were cut using a Leica UltraCut UCT (Leica Microsystems, Wetzlar, Germany). Semithin sections (1–2 μm) for LM were stained with toluidine blue and were examined with a LM Leitz Dialux 20 EB (Leica Microsystems). Ultrathin sections (800 Å) were stained with uranyl acetate replacement, and lead citrate (Electron Microscopy Sciences) and observed under a Zeiss EM 10 electron microscope (Zeiss, Oberkochen, Germany).
Blocks were cut using a Leica UltraCut UCT (Leica Microsystems, Wetzlar, Germany). Semithin sections (1–2 μm) for LM were stained with toluidine blue and were examined with a LM Leitz Dialux 20 EB (Leica Microsystems). Ultrathin sections (800 Å) were stained with uranyl acetate replacement, and lead citrate (Electron Microscopy Sciences) and observed under a Zeiss EM 10 electron microscope (Zeiss, Oberkochen, Germany).
4
Ultrastructural Analysis of Gill Tissue
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Gills were fixed for 4 h by immersion in 4% glutaraldehyde (Electron Microscopy Sciences, Hatfield, PA, USA) in phosphate-buffered saline (PBS 0.1 M, pH 7.2, 4 °C) and post-fixed for 2 h with osmium tetroxide (1% in PBS). After dehydration in graded ethanol, samples were placed in propylene oxide and embedded in Epon-Araldite (Araldite 502/Embed 812, Electron Microscopy Sciences, Hatfield, PA, USA). Samples were cut Using a Leica UltraCut UCT (Leica Microsystems, Wetzlar, Germany); semi-thin sections (1µm thickness) were stained with toluidine blue and observed using a LM Leitz Dialux 20 EB (Leica Microsystems, Wetzlar, Germany). Ultra-thin sections (800Å thickness) were stained with a uranyl acetate replacement, contrasted with lead citrate (Electron Microscopy Sciences, Hatfield, PA, USA), and finally observed under a Zeiss EM 10 electron microscope (Zeiss, Oberkochen, Germany).
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