The cells were fixed in 2.5% glutaraldehyde in 0.1M phosphate buffer pH 7.4 at room temperature for 1h followed by storage at +4 °C. Following the primary fixation the cells were rinsed with 0.1M phosphate buffer and Milli-Q® water before postfixed in 2% osmium tetroxide in 0.1M phosphate buffer, pH 7.4 at 4 °C for 2 hours. The cells were then stepwise ethanol dehydrated followed by stepwise acetone/LX-112 infiltration and finally embedded in LX-112 (Ladd Research). Ultrathin sections (approximately 60–80nm) were prepared using an EM UC 7 ultramicrotome (Leica Microsystems) and contrasted with uranyl acetate followed by Reynolds lead citrate and examined in a HT7700 transmission electron microscope (Hitachi High-Tech) operated at 100 kV. Digital images were acquired using a 2kx2k Veleta CCD camera (Olympus Soft Imaging Solutions GmbH).
2kx2k veleta ccd camera
Manufactured by Olympus
Sourced in Germany
The 2kx2k Veleta CCD camera is a high-resolution digital imaging device designed for laboratory and scientific applications. It features a 2048 x 2048 pixel CCD sensor, providing a resolution of approximately 4.2 megapixels. The camera is capable of capturing detailed images and data for various scientific and research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Ht7700 electron microscope, by Hitachi (6 mentions)
Ht7700 transmission electron microscope, by Hitachi (5 mentions)
Em uc7, by Leica (5 mentions)
Em uc6, by Leica (3 mentions)
400 mesh copper grid, by Ted Pella (3 mentions)
Ht7700, by Hitachi (3 mentions)
Em uc7 ultramicrotome, by Leica (2 mentions)
Em uc6 microtome, by Leica (2 mentions)
Lx112 resin, by Ladd Research Industries (1 mentions)
Lx 112, by Ladd Research Industries (1 mentions)
15 protocols using 2kx2k veleta ccd camera
1
Transmission Electron Microscopy Specimen Preparation
2
Ultrastructure Analysis of Testis Samples
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Testis piece from F1, F2 and F3 male offspring were fixed in 2.5% glutaraldehyde in 0.1 M phosphate buffer, pH 7.4 at room temperature for 30 min. Tissues were rinsed in 0.1 M phosphate buffer before post-fixation using 2% osmium tetroxide in 0.1 M phosphate buffer, pH 7.4 at 4°C for 2 h. The tissues were subsequently dehydrated in ethanol followed by acetone and finally embedded in LX-112. Ultrathin sections were prepared using a Leica EM UC7 (Leica Microsystems) and contrasted with uranyl acetate followed by Reynolds lead citrate. The sections were examined in a Hitachi HT 7700 Electron microscope (Hitachi High-Technologies) at 80kV and images acquired using a 2k x 2k Veleta CCD camera (Olympus Soft Imaging Solutions GmbH).
3
Ultrastructural Analysis of HeLa and MEF Cells
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Electron microscopy was performed on HeLa and MEF cells after 2 and 1 week recovery periods, respectively. After this period, cells were seeded on 6-well plates and the day after the cells were washed twice with 1xPBS and left with 0.25% trypsin for 5 min. Thereafter, the cells were washed with PBS twice with gentle centrifugation, and the supernatant was then removed and resuspended with 1 mL of fixative solution (2.5% glutaraldehyde in 0.1 M phosphate buffer, pH 7.4) for 1 h at room temperature and then store at 4°C before processing. The fixed cells were rinsed in 0.1 M phosphate buffer, pH 7.4 and post-fixed in 2% osmium tetroxide 0.1 M phosphate buffer, pH 7.4 at 4°C for 2 h, followed by stepwise ethanol and acetone dehydration and finally embedded in LX-112 (Ladd, 21,210 – LX 112). Ultrathin sections (approximately 60–80 nm) were prepared using a Leica EM UC 7 ultramicrotome (Leica, Vienna, Austria) and contrasted with uranyl acetate followed by lead citrate and imaged in a Hitachi HT7700 transmission electron microscope (Hitachi Hightech, Japan) at 100 kV. Digital images were acquired using a 2 kx2k Veleta CCD camera (Olympus Soft Imaging Solutions, GmbH, Münster, Germany).
4
Ultrastructural Analysis of Apoptotic Cells
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For ultrastructural analysis using TEM, Jurkat cells were triggered to undergo cell death for 24 h. Additionally, macrophages co-incubated with dying cells for 1 h were also processed for TEM. Cells were fixed in 2.5% glutaraldehyde in 0.1 M phosphate buffer, pH 7.4 at room temperature for 30 min and further fixed overnight in the refrigerator. The samples were rinsed in 0.1 M phosphate buffer and centrifuged prior to post-fixation using 2% osmium tetroxide in 0.1 M phosphate buffer, pH 7.4 at 4 °C for 2 h. Following the post-fixation, the cells were step-wise dehydrated in ethanol followed by acetone and LX-112 infiltration and finally embedded in LX-112. Ultrathin sections (approx. 50–60 nm) were prepared using a Leica EM UC6, contrasted with uranyl acetate followed by lead citrate, and examined in Hitachi HT 7700 electron microscope (Hitachi High-Technologies). Digital images were acquired using a 2kx2k Veleta CCD camera (Olympus Soft Imaging Solutions).
5
Ultrastructural Analysis of Dorsal Root Ganglia
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Mice were perfused with 2.5% glutaraldehyde and 1% formaldehyde in 0.1 M phosphate buffer, pH 7.4, followed by post-fixation in the same solution (>24 h). Following fixation, the DRGs were rinsed in 0.1 M phosphate buffer prior to post-fixation in 2% osmium tetroxide in 0.1 M phosphate buffer, pH 7.4, at 4°C for 2 h. DRGs were then stepwise dehydrated in ethanol followed by acetone and resin embedded in LX-112 (Ladd). Ultrathin sections (∼80–100 nm) were prepared using an EM UC 7 (Leica) and contrasted with uranyl acetate followed by lead citrate. The sections were examined using a Hitachi HT7700 transmission electron microscope (Hitachi High-Technologies) at 80 kV and digital images were acquired using a 2kx2k Veleta CCD camera (Olympus Soft Imaging Solutions).
6
Ultrastructural Analysis of Engrafted Liver Spheroids
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Whole eyes were dissected in cold PBS to obtain the engrafted liver spheroids and the surrounding iris. The fresh explant was fixed in 2.5% glutaraldehyde and 1% formaldehyde in 0.1 M phosphate buffer pH 7.4 for 1 h at RT followed by storage at 4 °C until further processing. After fixation, the explant was rinsed in 0.1 M phosphate buffer and treated with 2% osmium tetroxide in 0.1 M phosphate buffer pH 7.4 for 2 h at 4 °C. Following stepwise dehydration in ethanol and acetone, the graft was embedded in LX-112 resin (Ladd, Vermont, US). Ultrathin sections (approximately 80–100 nm) were prepared using an EM UC7 (Leica), contrasted with uranyl acetate, followed by lead citrate, and finally examined in an HT7700 transmission electron microscope (Hitachi High-Technologies, Tokyo, Japan) at 80 kV. Digital images were acquired using a 2kx2k Veleta CCD camera (Olympus Soft Imaging Solutions, Münster, Germany).
7
Ultrastructural Analysis of Liver Tissue
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Liver tissues were processed for ultrastructural analysis by transmission electron microscopy (TEM). Liver tissues were fixed in 2.5% glutaraldehyde in 0.1 M phosphate buffer (pH = 7.4) at room temperature for 30 min and further fixed overnight in the refrigerator. The samples were rinsed in 0.1 M phosphate buffer and centrifuged prior to postfixation using 2% osmium tetroxide in 0.1 M phosphate buffer (pH = 7.4) at 4 °C for 2 h. Following postfixation, the cells were dehydrated stepwise in ethanol followed by acetone and LX-112 infiltration and finally embedded in LX-112. Ultrathin sections (approximately 50–60 nm) were prepared using a Leica EM UC6, contrasted with uranyl acetate followed by lead citrate, and examined under a Hitachi HT 7700 electron microscope (Hitachi High-Technologies). Digital images were acquired using a 2kx2k Veleta CCD camera (Olympus Soft Imaging Solutions).
8
Silica NPs Uptake in THP-1 Cells
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THP-1 cells maintained in complete cell culture
medium at 1.0 × 106 cells per well were incubated
with 10 μg/mL silica NPs for 2 and 12 h. TEM samples were processed
according to the procedure described previously.64 (link) In brief, at the end of the incubation time, cells were
washed thrice with serum-free medium and fixed with 4% glutaraldehyde
in 0.1 M sodium phosphate buffer, pH 7.4 for 1 h at 4 °C. Following
postfixation in 1% OsO4 in 0.1 sodium phosphate buffer
for 1 h at 4 °C, the samples were serially dehydrated in a gradient
of ethanol followed by acetone and LX-112 infiltration and finally
embedded in LX-112. Ultrathin sections (approximately 50–80
nm) were prepared using a Leica EM UC6, contrasted with uranyl acetate
followed by lead citrate, and examined in Hitachi HT 7700 electron
microscope (Hitachi High-Technologies). Digital images were acquired
using a 2kx2k Veleta CCD camera (Olympus Soft Imaging Solutions).
medium at 1.0 × 106 cells per well were incubated
with 10 μg/mL silica NPs for 2 and 12 h. TEM samples were processed
according to the procedure described previously.64 (link) In brief, at the end of the incubation time, cells were
washed thrice with serum-free medium and fixed with 4% glutaraldehyde
in 0.1 M sodium phosphate buffer, pH 7.4 for 1 h at 4 °C. Following
postfixation in 1% OsO4 in 0.1 sodium phosphate buffer
for 1 h at 4 °C, the samples were serially dehydrated in a gradient
of ethanol followed by acetone and LX-112 infiltration and finally
embedded in LX-112. Ultrathin sections (approximately 50–80
nm) were prepared using a Leica EM UC6, contrasted with uranyl acetate
followed by lead citrate, and examined in Hitachi HT 7700 electron
microscope (Hitachi High-Technologies). Digital images were acquired
using a 2kx2k Veleta CCD camera (Olympus Soft Imaging Solutions).
9
Ultrastructural Analysis of Cercariae
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The cercariae specimens from 96% ethanol were rinsed in 0.1M phosphate buffer prior to postfixation in 2% osmium tetroxide in 0.1M phosphate buffer, pH 7.4 at 4 C for 2 h. Following stepwise dehydration in ethanol and acetone, the specimens were embedded in LX-112 resin (Ladd). Ultrathin sections (approximately 80-100 nm) prepared using a EM UC7 (Leica) and contrasted with uranyl acetate followed by lead citrate were examined in a HT7700 transmission electron microscope (Hitachi High-Technologies,) at 80 kV. Digital images were acquired using a 2kx2k Veleta CCD camera (Olympus Soft Imaging Solutions).
10
Transmission Electron Microscopy of Nanomaterial Uptake
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TEM imaging was performed on D. magna samples after exposure to COOH-PS NPs or NH2-PS NPs. The samples were prefixed with 4% glutaraldehyde in 0.1 M sodium phosphate buffer pH 7.4 for 1 h at 4 °C. After that they were fixed in 1% OsO4 in 0.1 sodium phosphate buffer for 1 h at 4 °C and were subsequently dehydrated using a gradient of ethanol followed by acetone and LX-112 infiltration followed by embedding in LX-112 resin (Ladd Research Industries, Vermont, OH). Ultrathin sections (50–80 nm) were prepared using a Leica EM UC6 microtome and these were further contrasted with uranyl acetate followed by lead citrate, and finally examined using a Hitachi HT 7700 electron microscope (Hitachi High-Technologies). We used the 2kx2k Veleta CCD camera (Olympus) for image acquisition. TEM analysis was also performed on HT-29 cells. Briefly, cells were detached from the transwells after cell culture and prefixed with 4% glutaraldehyde in 0.1 M sodium phosphate buffer pH 7.4 for 1 h at 4 °C. Thereafter, the procedure described for D. magna samples was followed.
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