Staphylococcus aureus FtsZ (12 μM) was incubated in the absence and in the presence of 3 μg/mL of 1 in 50 mM MOPS buffer (pH 6.5) at 25°C. After 10 min, 5 mM MgCl2, 50 mM KCl, and 1 mM GTP were added to the reaction mixtures and incubated at 37°C for 15 min. Then, 10 μL of the sample mixtures were placed on a glow-discharged Formvar carbon-coated copper grid (400 mesh) for 10 min. The grids were subsequently subjected to negative staining using 10 μL of 0.5% phosphotungstic acid (PTA) for 30 s, air-dried and digital images of the specimen were observed with a transmission electron microscope (JEOL model JEM 2010) operated at 200 kV and equipped with a Gatan MSC 794 CCD camera.
Msc 794 ccd camera
Manufactured by Ametek
The MSC 794 CCD camera is a high-performance imaging device designed for scientific and industrial applications. It features a 7.4 megapixel CCD sensor with a pixel size of 7.4 μm x 7.4 μm, providing excellent image quality and resolution. The camera offers a range of connectivity options, including USB 3.0 and GigE interfaces, enabling seamless integration with various systems and software platforms.
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4 protocols using msc 794 ccd camera
1
Staphylococcus aureus FtsZ Polymerization Inhibitor
2
Visualizing Drug Aggregate Formation
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TEM was used to examine the formation of ‘drug aggregates’ by small molecules under aqueous conditions. Samples of 10 μM Congo red (dissolved in deionized water) and TIPP (dissolved in 0.5% DMSO) were incubated with the labelled V216C mutant (20 nM). A drop of the solution mixture (10 μl) was then applied onto a carbon grid (400 mesh), stained with 0.5% PTA (phosphotungstic acid) and then dried overnight. TEM pictures were obtained with a 200 kV DEOL JEM-2010 electron microscope equipped with a Gatan MSC 794 CCD camera.
3
FtsZ Polymerization Visualization Protocol
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S. aureus FtsZ (12 µM) was incubated in the absence and in the presence of different concentrations of the test compounds in 50 mM MOPS buffer (pH 6.5) at 25°C. After 10 min, 5 mM MgCl2, 50 mM KCl, and 1 mM GTP were added to the reaction mixtures and incubated at 37°C for 15min. Then, 10 µL of the sample mixtures were placed on a glow-discharged Formvar carbon-coated copper grid (400 mesh) for 10 min. The grids were subsequently subjected to negative staining using 10 µL of 0.5% phosphotungstic acid (PTA) for 30s, air-dried and digital images of the specimen were observed with a transmission electron microscope (JEOL model JEM 2010) operated at 200 kV and equipped with a Gatan MSC 794 CCD camera.
+ Expand
S. aureus FtsZ (12 µM) was incubated in the absence and in the presence of different concentrations of the test compounds in 50 mM MOPS buffer (pH 6.5) at 25°C. After 10 min, 5 mM MgCl2, 50 mM KCl, and 1 mM GTP were added to the reaction mixtures and incubated at 37°C for 15min. Then, 10 µL of the sample mixtures were placed on a glow-discharged Formvar carbon-coated copper grid (400 mesh) for 10 min. The grids were subsequently subjected to negative staining using 10 µL of 0.5% phosphotungstic acid (PTA) for 30s, air-dried and digital images of the specimen were observed with a transmission electron microscope (JEOL model JEM 2010) operated at 200 kV and equipped with a Gatan MSC 794 CCD camera.
4
Characterization of AuNP Assemblies
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The assembled structures were deposited on freshly cleaned muscovite mica (Electron Microscopy Sciences) for five minutes, then rinsed with MQ-pure water and dried under nitrogen gas. AFM imaging was performed in air in ScanAsyst mode on a Veeco Multimode Scanning Probe Microscope with silicon Scanasyst-air probes (resonant frequency 50–90 kHz, spring constant 0.4 N m−1, and a tip radius of 2 nm).
For TEM imaging, a JEM 2010 transmission electron microscope (JEOL) operated at 200 kV was used. The images were analyzed using a GATAN MSC 794 CCD Camera and GATAN Digital Microscopy software. Five μL AuNP assemblies were transferred to SPI® Supplies Holey carbon coated grids onto 400 mesh copper, incubated for three minutes, washed with 50 μL of MQ-pure water, and dried under nitrogen gas. Samples were then dried completely for at least 15 hours under ambient conditions in a vacuum desiccator.
For TEM imaging, a JEM 2010 transmission electron microscope (JEOL) operated at 200 kV was used. The images were analyzed using a GATAN MSC 794 CCD Camera and GATAN Digital Microscopy software. Five μL AuNP assemblies were transferred to SPI® Supplies Holey carbon coated grids onto 400 mesh copper, incubated for three minutes, washed with 50 μL of MQ-pure water, and dried under nitrogen gas. Samples were then dried completely for at least 15 hours under ambient conditions in a vacuum desiccator.
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