Samples were processed in the Electron Microscopy Core at Fudan Univeristy. Cell pellets were fixed with 2.5% glutaraldehyde in 0.1 M phosphate buffer. The cells were washed with 0.1 M sodium cacodylate buffer and postfixed with 1% osmium tetroxide. The pellets were then dehydrated in graded ethanol series, infiltrated, and embedded in Spurr's resin. Samples were then polymerized for 48 h at 60°C, cut into 60-nm-thick sections on LKB-I microtome, positioned on 200 mesh grids, and stained with uranyl acetate and lead citrate. TEM was performed on PHILIPS CM120 TEM at an accelerating voltage of 120 Kv. Images were acquired with Gatan type UltraScan 4000SP CCD Camera connected to the TEM.
Ultrascan 4000sp ccd camera
Manufactured by Ametek
The UltraScan 4000SP CCD Camera is a high-performance scientific imaging device designed for laboratory applications. It features a large-format CCD sensor that captures detailed and high-resolution images. The camera offers exceptional sensitivity, low noise, and fast readout speeds to support a wide range of scientific imaging tasks.
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7 protocols using ultrascan 4000sp ccd camera
1
Transmission Electron Microscopy of Cells
2
Electron Microscopy Sample Preparation Protocol
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Cell samples were processed by the Electron Microscopy Core at Fudan University39 (link). Cell pellets were fixed with 2.5% glutaraldehyde in 0.1 M phosphate buffer and then washed with 0.1 M sodium cacodylate buffer and post fixed with 1% osmium tetroxide. The pellets were dehydrated in a graded ethanol series, infiltrated, and embedded in Spurr’s resin. Samples were polymerized for 48 h at 60 °C, cut into 60-nm-thick sections using an LKB-I microtome, positioned on 200-mesh grids, and stained with uranyl acetate and lead citrate. TEM was performed using a PHILIPS CM120 TEM at an accelerating voltage of 120 kV. Images were acquired with a connected Gatan type Ultra Scan 4000SP CCD camera.
3
Transmission Electron Microscopy of SFD Vaccine Formulations
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A Philips CM120 transmission electron microscope was used to make transmission electron microscopy images. SFD powders containing WIV (5 µg HA formulation) with or without adjuvants i.e. BLP (300 µg in 1 mg of SFD powder), Advax (500 µg in 1 mg of SFD powder) were reconstituted in sterile filtered water. Liquid and reconstituted SFD formulations were placed on a plain carbon grid, rinsed with water and then samples were stained twice with 5 µL of 2 wt-% uranyl acetate. A Gatan type UltraScan 4000SP CCD Camera at a magnification was used to take images.
4
Ultrastructural Analysis of Cellular Samples
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Samples were processed in the Electron Microscopy Department at Shanghai Jiao Tong University. Cell pellets were fixed with 2.5% glutaraldehyde overnight in PBS and post-fixed with 1% osmium tetroxide (pH 7.4) for 2 h at room temperature. The pellets were then dehydrated in a graded ethanol series and infiltrated with Spurr’s resin to embed the tissues. Samples were then polymerized for 48 h at 60 °C, cut into 60-nm-thick sections on an LKB-I microtome (LKB, Sweden), positioned on 200-mesh grids and stained with uranyl acetate and lead citrate. TEM was performed on a PHILIPS CM120 transmission electron microscope at an accelerating voltage of 120 kV. Images were acquired with a Gatan-type UltraScan 4000SP CCD camera (Pleasanton, CA) connected to the microscope.
5
Transmission Electron Microscopy Protocol
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Samples were processed in the department of pathology at Jinling hospital. Cell pellets were xed with 2.5% glutaraldehyde in 0.1 M phosphate buffer. The cells were washed with 0.1 M sodium cacodylate buffer and post xed with 1% osmium tetroxide. The pellets were then dehydrated in graded ethanol series, in ltrated, and embedded in Spurr's resin. Samples were then polymerized for 48 h at 60°C, cut into 60-nm-thick sections on LKB-I microtome, positioned on 200 mesh grids, and stained with uranyl acetate and lead citrate. The ultrathin sections were mounted on JEOL JEM-1011 TEM at an accelerating voltage of 120 Kv. Images were acquired with Gatan type UltraScan 4000SP CCD Camera connected to the TEM. The numbers of autophagosome were calculated manually for a minimum of 50 cells for each sample and the photos were taken.
6
Ultrastructural Analysis of MMQ Xenograft Tumors
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Samples were tested at the Electron Microscopy Center in Shanghai Jiaotong University. The tumor tissue obtained from the MMQ xenograft tumors of mice were fixed with 2.5% glutaraldehyde overnight and post-fixed with 1% osmium tetroxide (pH 7.4) for 2 h at room temperature. The pellets were then dehydrated in graded ethanol series, infiltrated and embedded in Spurr's resin (01916-1; Polysciences). Samples were then polymerized for 48 h at 60°C, cut into 60-nm-thick sections on LKB-I microtome (LKB, Sweden), positioned on 200-mesh grids (22896-1; Polysciences) and stained with uranyl acetate and lead citrate. TEM was performed on PHILIPS CM120 TEM at an accelerating voltage of 120 kV. Furthermore, images were acquired using Gatan type UltraScan 4000SP CCD Camera connected to the transmission electron microscope.
7
Transmission Electron Microscopy Protocol
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Samples were processed in the department of pathology at Jinling hospital. Cell pellets were xed with 2.5% glutaraldehyde in 0.1 M phosphate buffer. The cells were washed with 0.1 M sodium cacodylate buffer and post xed with 1% osmium tetroxide. The pellets were then dehydrated in graded ethanol series, in ltrated, and embedded in Spurr's resin. Samples were then polymerized for 48 h at 60°C, cut into 60-nm-thick sections on LKB-I microtome, positioned on 200 mesh grids, and stained with uranyl acetate and lead citrate. The ultrathin sections were mounted on JEOL JEM-1011 TEM at an accelerating voltage of 120 Kv. Images were acquired with Gatan type UltraScan 4000SP CCD Camera connected to the TEM. The numbers of autophagosome were calculated manually for a minimum of 50 cells for each sample and the photos were taken.
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