Briefly, cells were washed 3x with 0.1 M sodium cacodylate buffer, post-fixed in 1% osmium tetroxide (2 h), rinsed with dH2O, and dehydrated in a graded ethanol series (50–100%). Samples were en bloc stained with uranyl acetate in the 70% ethanol step for 30 min at 4 °C. Following dehydration, cells were infiltrated with resin (Agar 100)/ethanol mixtures. The next day, cells were infiltrated and embedded with 100% epoxy resin in inverted BEEM-capsules for two days (60 °C). Ultrathin sections of 50 nm thickness were cut following the separation of polymerized cells from the glass bottom dishes (via freeze-thaw approach) and post-stained with 3% uranyl acetate in water (10 min) and Reynold’s lead citrate (2 min). Micrographs were taken in a JEOL JEM1400 (JEOL, Japan) equipped with an Olympus SIS Quemesa 11 Mpxl camera at 80 KV.
Sis quemesa 11 mpxl camera
Manufactured by Olympus
Sourced in Japan, Belgium
The SIS Quemesa 11 Mpxl camera is a digital imaging device designed for laboratory applications. It features an 11-megapixel sensor and captures high-resolution images. The core function of this camera is to provide reliable and accurate image data for scientific and research purposes.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using sis quemesa 11 mpxl camera
1
Ultrastructural Analysis of Cells via TEM
2
Ultrastructural Analysis of Organoids
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Organoids were fixed overnight in 2.5% glutaraldehyde (Grade I, Sigma-Aldrich) in 0.1 M sodium cacodylate buffer (pH 7.2) at 4°C. The fixed organoids were pelleted at 200 g and washed three times in 0.1 M cacodylate buffer.
The pellets were resuspended in 2.0% low melting point agarose (Thermo Fisher Scientific) and centrifuged at 1000 g. After solidification, the pellets were cut up in small cubes, washed and post-fixed in 1% osmium tetroxide and 1.5% potassium ferrocyanide in 0.1 M cacodylate buffer, washed again and mordanted by 0.1% tannic acid for 20 min. Then, samples were stained overnight en bloc with uranyl acetate (Agar Scientific, Stansted, UK) followed by 30-min lead aspartate incubation with intermittent washing in cacodylate buffer, dehydrated in a graded ethanol series and propylene oxide and finally embedded in epoxy resin (Agar 100; Agar Scientific). The embedded samples were polymerized for 2 days at 60°C. Thin 70 nm sections were cut with a UCT ultramicrotome (Leica) and analyzed using the JEM1400 transmission electron microscope (JEOL, Zaventem, Belgium) equipped with an Olympus SIS Quemesa 11Mpxl camera.
The pellets were resuspended in 2.0% low melting point agarose (Thermo Fisher Scientific) and centrifuged at 1000 g. After solidification, the pellets were cut up in small cubes, washed and post-fixed in 1% osmium tetroxide and 1.5% potassium ferrocyanide in 0.1 M cacodylate buffer, washed again and mordanted by 0.1% tannic acid for 20 min. Then, samples were stained overnight en bloc with uranyl acetate (Agar Scientific, Stansted, UK) followed by 30-min lead aspartate incubation with intermittent washing in cacodylate buffer, dehydrated in a graded ethanol series and propylene oxide and finally embedded in epoxy resin (Agar 100; Agar Scientific). The embedded samples were polymerized for 2 days at 60°C. Thin 70 nm sections were cut with a UCT ultramicrotome (Leica) and analyzed using the JEM1400 transmission electron microscope (JEOL, Zaventem, Belgium) equipped with an Olympus SIS Quemesa 11Mpxl camera.
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