For scanning electron microscopy, specimens were initially prepared by fixing in 2.5% glutaraldehyde in 0.15 M sodium phosphate buffer, pH 7.4, at 4 °C for 24–48 h. Post-fixation was carried out in 1% buffered osmium tetroxide overnight at 4 °C, then samples were dehydrated through a graded ethanol series, followed by critical point drying using a BioRad E3000 critical point dryer (Quorum Technologies, East Sussex, England), or by dehydration with acetone and hexamethyldisilazane. All samples prepared for SEM were sputter coated with 10 nm gold using a Hummer VII (Anatech USA, Union City, California, USA). Micrographs were taken using a JEOL JSM-6500F Field Emission Scanning Electron Microscope at the Central Instrument Facility, Imaging Laboratory, located at Colorado State University. All images were captured digitally as tiff files, and graphically edited to input black image backgrounds (Adobe Photoshop, CS6 13.0.1). All measurements were taken at the widest or longest point of a given structure (Table 1 ), and are given in μm. Ranges are presented, followed by the means in parentheses.
E3000 critical point dryer
Manufactured by Quorum Technologies
Sourced in United Kingdom
The E3000 critical point dryer is a laboratory equipment designed for the drying of samples in a controlled environment. It functions by transitioning a sample's liquid phase directly to the gas phase, avoiding the surface tension effects associated with conventional drying methods.
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Lab products found in correlation
2 protocols using e3000 critical point dryer
1
Scanning Electron Microscopy Sample Preparation
2
Preparation of Cells for SEM Imaging
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Cells were removed from their media by centrifugation at 2000 rpm for 5 min and washed and resuspended in PBS buffer. The washing process was done three times to remove access media peptone. The cells were then fixed in a 1 wt% glutaraldehyde PBS buffer solution for 1 hour at room temperature, the cells were then centrifuged at 2000 rpm and the pellet washed with deionized water three times to remove excess glutaraldehyde, the cell samples were then re-suspended in and dehydrated 50%/75%/90% and absolute ethanol solutions for 30 minutes per each ethanol concentration. Cells were then swabbed onto a glass slide and submerged into absolute ethanol and dried using liquid CO2 at its critical point using an E3000 Critical Point Dryer (Quorum Technologies, UK) and then coated in 10 nm Carbon. The samples were imaged using an Ultra-High-Resolution Scanning Electron Microscope using cold field emission (Hitachi SU8230, Japan).
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