Samples (APCs or telomere vesicles purified by fluorescence-activated vesicle sorting), were adhered on Ply-L-Lysine-coated round glass coverslips of 1 cm diameter, then fixed with 2.5% glutaraldehyde in 0.1M sodium cacodylate buffer (pH 7.4) at room temperature for 1h. Fixed samples were rinsed 10 minutes with 0.1M sodium cacodylate buffer (3 times), then dehydrated through ethanol gradients (30%, 50%, 70%, 85%, 95%, 100% - 10 mins at room temperature for each ethanol concentration). A 1:1 ethanol: hexamethyldisilazane (HMDS) mix was then added to the samples for 5 min at room temperature followed by a final 5 min step incubation in HMDS. All samples were then left drying overnight. The upper base of each aluminium stub was coated with colloidal silver paint and the coverslips were pasted onto them. Samples mounted on stubs were coated with a gold layer by Q150R S Rotary-Pumped Sputter Coater (Quorum Technologies) and examined by FESEM (Sigma-Zeiss) at an accelerating voltage of 2 kV using secondary electron (SE) detection. A number of regions of interest (ROIs) containing single or multiple APCs for each sample were recorded to confirm absence of glass beads or to depict structural alterations. Micrographs of telomere vesicles were used to perform particle size distribution analysis by measuring the diameter of each adhered telomere vesicle.
Q150r s rotary pumped sputter coater
Manufactured by Quorum Technologies
Sourced in United Kingdom
The Q150R S Rotary-Pumped Sputter Coater is a laboratory equipment designed for depositing thin films on samples. It utilizes a rotary pump to create a vacuum environment necessary for the sputtering process. The core function of this product is to coat samples with thin layers of materials, which can be useful in various scientific and industrial applications.
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Lab products found in correlation
3 protocols using q150r s rotary pumped sputter coater
1
Scanning Electron Microscopy of APCs and Telomere Vesicles
2
Scanning Electron Microscopy of APCs and Telomere Vesicles
3
SEM Imaging of Neuronal Cultures
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Neuronal cultures where incubated with PBS, human pooled serum (HPS), heat-inactivated MMN serum, MMN serum, or heat-inactivated MMN serum with HPS as an external complement source. Incubation was performed at 378C for 15 or 30 minutes. Samples were fixed using 4% paraformaldehyde as described above. Serial dehydration was achieved by consecutive incubation steps in 12.5% EtOH in PBS, 25% EtOH in PBS, 50% EtOH in PBS, 75% EtOH in H 2 O, 90% EtOH in H 2 O, 100% EtOH, followed by incubation in 50% EtOH/50% hexamethyldisilazane (HDMS) and finally 100% HDMS. Samples were mounted onto aluminum specimen mounts (Agar Scientific, Stansted, UK), followed by coating with 1nm gold using a Q150R S Rotary-Pumped Sputter Coater (Quorum Technologies, Lewes, UK). Samples were examined with a Nova Nano-SEM 200 scanning electron microscope (FEI, Hillsboro, OR) operated with an accelerated voltage of 10kV at a magnification of 37,500 using a Phenom Pro desktop scanning electron microscope and Pro Suite software (Phenom-World, Eindhoven, the Netherlands).
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