SEM images of E. coli on 80% lean ground beef were taken using an ultra-high resolution field emission scanning electron microscope (Hitachi, Schaumburg, IL, USA; model S-4800 Type II). 100 μL of water (negative control) or E. coli solution (105 or 108 CFU/mL) was applied to 1.5 g of thawed ground beef in a petri dish and mixed on a nutator mixer for 30 minutes. After mixing, the fixative was added to fix the sample overnight. The fixative consisted of 4% v/v paraformaldehyde and 1% v/v glutaraldehyde with phosphate buffer. The fixed samples were then dried in a Polaron Critical Point Dryer (Quorum Technologies, East Sussex, UK; model E3000) and platinum-coated using a Hammer 6.2 Sputter Coater (Anatech, Union City, CA, USA) before SEM imaging.
Polaron critical point dryer
Manufactured by Quorum Technologies
Sourced in United Kingdom
The Polaron Critical Point Dryer is a laboratory equipment designed to dry samples while preserving their delicate structures. It utilizes the critical point drying process to remove liquid from samples without causing damage or deformation.
Automatically generated - may contain errors
4 protocols using polaron critical point dryer
1
SEM Imaging of E. coli on Ground Beef
2
Platelet Ultrastructure Analysis by SEM
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Platelets were fixed with 2.5% glutaraldehyde in PBS for 20 min, followed by post-fixation with 1% osmium tetroxide (OsO4) in PBS for 20 min. Next, platelets were dehydrated in graded aqueous ethanol solutions from 30% to 96% (each for 5 min), and then in 100% ethanol (three steps of 5 min each). Samples were dried in Polaron Critical Point Dryer (Quorum Technologies Ltd., Kent, UK) and coated with a thin layer of gold in sputter coater images were acquired in Zeiss Supra 40 VP SEM.
3
Scanning Electron Microscopy of Micranthemum Capitula
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SEM was performed on M. inodora capitula at different developmental stages. Tissue was fixed in the fixation solution (4% [w/v] PFA, 0.01% [v/v] DMSO, 1× PBS, 0.001% [v/v] TWEEN 20, and 0.001% [v/v] Triton X-100) overnight at 4°C. The next day, tissues were dehydrated in an ethanol series (30% [v/v], 50% [v/v], 70% [v/v], 85% [v/v], 95% [v/v], 100% [v/v]) for 30 min per step. Dehydrated tissues then underwent critical point drying using a Polaron critical point dryer (Quorum Technologies) and were mounted onto SEM stubs (Agar Scientific) using carbon tape (Agar Scientific). Mounted stubs were sputter-coated with gold for 2 min using a Polaron E5100 sputter-coater (Quorum Technologies). Samples were then imaged on a Quanta 250 FEG (FEI UK) using the secondary detector.
4
Scanning Electron Microscopy Sample Preparation
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The dissected parts were dried in an ascending ethanol series (70%, 80%, 90%, and 100% (twice)) followed by critical point drying in CO2 (Polaron Critical Point Dryer, Quorum Technologies, Laughton, UK). They were then cleaned by being submerged in 30% hydrogen peroxide solution for 5 min. The objects were dehydrated again for 5 min in 100% ethanol.
The specimens were mounted on a roof-like structure of silver tape attached to a standard SEM object mount. They were stabilized using conductive silver paint (all materials from Plano GmbH, Wetzlar, Germany). Objects were gold-coated for 5 min from various angles to improve electrical properties (Quorum Emitech K550X, Ashford, UK).
Images were taken using a Scanning Electron Microscope (ZEISS EVO LS 10, Oberkochen, Germany) by using the secondary electron detector and, if necessary, the backscatter detector. An acceleration voltage of 15 kV was used.
The terminology and method for mouthpart descriptions follows those of [26 (link)].
The specimens were mounted on a roof-like structure of silver tape attached to a standard SEM object mount. They were stabilized using conductive silver paint (all materials from Plano GmbH, Wetzlar, Germany). Objects were gold-coated for 5 min from various angles to improve electrical properties (Quorum Emitech K550X, Ashford, UK).
Images were taken using a Scanning Electron Microscope (ZEISS EVO LS 10, Oberkochen, Germany) by using the secondary electron detector and, if necessary, the backscatter detector. An acceleration voltage of 15 kV was used.
The terminology and method for mouthpart descriptions follows those of [26 (link)].
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