Cells were incubated at 37°C or 45°C for 2 hours with or without vancomycin at MIC level, then washed once with 0.1 M Sorensen’s Buffer and resuspended in 2.5% glutaraldehyde. These were embedded in EMbed-812 resin (Electron Microscopy Sciences, Hatfield PA). Ultra-thin sections, 70 nm in thickness, were mounted on copper grids for transmission electron microscopy (TEM). Grids were post-stained with uranyl acetate and Reynolds lead citrate. Sections were examined using a Philips CM100 Transmission Electron Microscope at 60 kV. Images were recorded digitally using a Hamamatsu ORCA-HR digital camera system, operated using AMT software (Advanced Microscopy Techniques Corp., Danvers, MA). The cell wall thickness was determined in ImageJ by identifying the geometric center of each cell and constructing a radius through a representative region of cell wall. The length of radius passing from the cell membrane to the outer edge of the cell wall was defined as the cell wall thickness.
Cm 100 transmission electron microscope
Manufactured by Hamamatsu Photonics
Sourced in United States
The CM 100 transmission electron microscope is a laboratory equipment designed for high-resolution imaging of samples. It utilizes a focused beam of electrons to magnify and project an image of the internal structure of specimens onto a fluorescent screen or digital sensor. The core function of the CM 100 is to provide detailed visualization and analysis of microscopic features within materials, cells, or other samples.
Automatically generated - may contain errors
3 protocols using cm 100 transmission electron microscope
1
Quantifying Cell Wall Thickness via TEM
2
Ultrastructural Analysis of Transverse Sinus in Mice
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After 24 h of IVH, the mice were anesthetized and subsequently subjected to intracardiac perfusion with a solution of 0.1 mol/L Sorensen's buffer (pH 7.4) containing 4% paraformaldehyde and 2.5% glutaraldehyde. The animals were dissected at 4 °C to retrieve the meninges. The meninges surrounding the transverse sinus (TS) were removed with microscissors, and the TS was sectioned and trimmed into small tissue blocks (1 mm3). Subsequently, the samples were fixed with a 1.0% OsO4 solution and dehydrated using a series of ethyl alcohol solutions with increasing concentrations. Once dehydration was complete, the specimens were infused with propylene oxide, embedded in Epon resin, and then sectioned. The stained samples were analyzed with a Philips CM 100 transmission electron microscope (Hillsboro, OR, USA) and digitally documented with a Hamamatsu ORCA-HR camera (Hamamatsu City, Shizuoka, Japan) 45 (link).
3
Electron Microscopy of Rat Subventricular Zone
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Electron microscopy was performed as previously described [26 (link)]. Rats were anaesthetised and subjected to intracardiac perfusion with 4% paraformaldehyde and 2.5% glutaraldehyde in 0.1 mol/l Sorensen's buffer (pH 7.4). The subventricular zones were removed from the brain, and a 1 mm thick coronal tissue slice was cut with a blade 4 mm overnight at 4°C. Samples were then postfixed with 1.0% OsO4 and dehydrated in graded ethyl alcohol. After dehydration was complete, the samples were infiltrated with propylene oxide, embedded in Epon resin and sectioned. Ultrathin sections were then stained with uranyl acetate and Reynold's lead citrate. Sections were evaluated using a Philips CM 100 transmission electron microscope (Hillsboro, OR, USA) and were digitally acquired using a Hamamatsu ORCA-HR camera (Hamamatsu City, Shizuoka, Japan).
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