The 3-d-old w1118 male reproductive tracts were dissected and incubated overnight in 2.5% glutaraldehyde and 4% formaldehyde in PBS (pH 7.2). Glands were then washed with PBS, refixed in 1% osmium tetroxide (Agar Scientific) for 20 min, washed three times in distilled water, and dehydrated through a graded alcohol series and incubated in ethanol and Spurr’s epoxy resin (1:1) (Agar Scientific). Glands were embedded in 100% Spurr’s epoxy resin between two sheets of polythene and polymerized overnight at 60 °C. Ultrathin sections were prepared with a Reichert Ultracut R Ultramicrotome (Leica Biosystems) and mounted on formvar-coated slot grids (Agar Scientific). Sections were contrasted with 2% uranyl acetate and lead citrate (Agar Scientific) and imaged using a JEOL 1010 electron microscope (80 kV).
Spurr s epoxy resin
Manufactured by Agar Scientific
Spurr's epoxy resin is a widely used embedding medium for the preparation of biological samples for electron microscopy. It is a low-viscosity, high-quality epoxy resin formulation designed to provide excellent sectioning properties and preservation of ultrastructural details.
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2 protocols using spurr s epoxy resin
1
Electron Microscopy of Drosophila Male Reproductive Tracts
2
Ultrastructural Analysis of Drosophila Reproductive Tracts
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The copyright holder for this preprint this version posted April 25, 2020. ; https://doi.org/10.1101/2020.04.24.059238 doi: bioRxiv preprint 3-day-old w 1118 male reproductive tracts were dissected and incubated overnight in 2.5% glutaraldehyde and 4% formaldehyde in PBS (pH 7.2). Glands were then washed with PBS, refixed in 1% osmium tetroxide (Agar Scientific) for 20 minutes, washed 3 times in distilled water and dehydrated through a graded alcohol series and incubated in ethanol and Spurr's epoxy resin (1:1) (Agar Scientific). Glands were embedded in 100% Spurr's epoxy resin between two sheets of polythene and polymerised overnight at 60°C. Ultrathin sections were prepared with a Reichert Ultracut R Ultramicrotome (Leica Biosystems) and mounted on formvar-coated slot grids (Agar Scientific). Sections were stained with 2% uranyl acetate and lead citrate (Agar Scientific), and imaged using a JEOL 1010 electron microscope (80kV).
The copyright holder for this preprint this version posted April 25, 2020. ; https://doi.org/10.1101/2020.04.24.059238 doi: bioRxiv preprint 3-day-old w 1118 male reproductive tracts were dissected and incubated overnight in 2.5% glutaraldehyde and 4% formaldehyde in PBS (pH 7.2). Glands were then washed with PBS, refixed in 1% osmium tetroxide (Agar Scientific) for 20 minutes, washed 3 times in distilled water and dehydrated through a graded alcohol series and incubated in ethanol and Spurr's epoxy resin (1:1) (Agar Scientific). Glands were embedded in 100% Spurr's epoxy resin between two sheets of polythene and polymerised overnight at 60°C. Ultrathin sections were prepared with a Reichert Ultracut R Ultramicrotome (Leica Biosystems) and mounted on formvar-coated slot grids (Agar Scientific). Sections were stained with 2% uranyl acetate and lead citrate (Agar Scientific), and imaged using a JEOL 1010 electron microscope (80kV).
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