In order to assess whether T. pseudonana was present on the coral surface, 9 nubbins for each experimental group (A and B) were treated as follows: samples were dehydrated by immersion in alcohol at increasing gradations (10, 30, 50, 70, 80, 90, 95 and 100%) and processed through critical point drying (Polaron CPD7501, Quorum Technologies, Newhaven, UK). Then, each coral piece was placed on a biadhesive tape mounted on a stub and sputtered with gold-palladium (Polaron SC7640 Sputter Coater, Quorum Technologies, Newhaven, UK) for observation under a field emission scanning electron microscopy FE SEM (Zeiss Supra 40, Carl Zeiss, Oberkochen, Germany).
Polaron cpd 7501
Manufactured by Quorum Technologies
Sourced in United Kingdom
The Polaron CPD 7501 is a critical point dryer for sample preparation in scanning electron microscopy (SEM) and transmission electron microscopy (TEM) applications. The device facilitates the drying of samples while maintaining their structural integrity.
Automatically generated - may contain errors
Lab products found in correlation
G5882, by Merck Group (2 mentions)
P4417, by Merck Group (2 mentions)
Polaron sc7640 sputter coater, by Quorum Technologies (1 mentions)
Supra 40, by Zeiss (1 mentions)
Carbohydrazide, by Merck Group (1 mentions)
Hexylene glycol, by Merck Group (1 mentions)
Quanta 200 sem, by Thermo Fisher Scientific (1 mentions)
Osmium tetroxide, by Merck Group (1 mentions)
Mc190 hd, by Leica (1 mentions)
Tl led, by Leica (1 mentions)
Quanta 250 feg sem, by Thermo Fisher Scientific (1 mentions)
Em ace 600 high vacuum sputterer, by Leica (1 mentions)
Fei quanta 200, by Thermo Fisher Scientific (1 mentions)
Jee 4c, by JEOL (1 mentions)
5 protocols using polaron cpd 7501
1
Scanning Electron Microscopy of Coral
2
Morphological Analysis of Cells on ND Nanofilm
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To assess the morphological changes in cells grown on 6-well plates uncoated and coated with ND nanofilm, scanning electron microscopy (SEM) was used (Quanta 200). Cells were maintained on nfND for 7 days without medium changes, then rinsed in phosphate-buffered saline (PBS, pH 7.2) and fixed in 2.5% glutaraldehyde (G5882, Sigma-Aldrich). Subsequent steps were performed using osmium tetroxide and carbohydrazide. Contrasted cells were dehydrated through a hexylene glycol series (Sigma-Aldrich) using a point dryer (Polaron CPD 7501, Quorum Technologies, Laughton, UK). Finally, samples were placed on aluminium SEM, and four independent images were taken for each sample.
3
Cellular Characterization of Primary Cell Types Using Optical and Scanning Electron Microscopy
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Primary mesenchymal cells originating from a hind limb (PMC), primary heart cells (PHC), primary neuronal cells originating from the brain (PNC), primary eye cells (PEC), and primary blood vessel cells isolated from the chorioallantoic membrane (PVC) were assessed using an optical inverted microscope (TL-LED, Leica Microsystems GmbH, Wetzlar, Germany) connected to a digital camera (Leica MC190 HD) using LAS V4.10 software. Cells were seeded (1 × 105 cells per well) in 35 mm diameter Petri dishes with and without GO scaffolds. After incubation for 24 h, samples were imaged.
PMC were additionally visualized using a Quanta 200 SEM (FEI, Hillsboro, OR, USA). Cells were seeded (1 × 105 cells per well) in 35 mm diameter Petri dishes with and without GO scaffolds. After a 96-h incubation period, samples were prepared as described by Heckman et al. [46 (link)]. Cells were fixed using 2.5% glutaraldehyde in phosphate-buffered saline (PBS) at pH 7.2, contrasted with 1% osmium tetroxide (Sigma-Aldrich) and 1% carbohydrazide (Sigma-Aldrich). Subsequently, cells were dehydrated in increasing concentrations of hexylene glycol (Sigma-Aldrich). Drying was performed using a Polaron CPD 7501 critical point dryer (Quorum Technologies, Laughton, UK).
PMC were additionally visualized using a Quanta 200 SEM (FEI, Hillsboro, OR, USA). Cells were seeded (1 × 105 cells per well) in 35 mm diameter Petri dishes with and without GO scaffolds. After a 96-h incubation period, samples were prepared as described by Heckman et al. [46 (link)]. Cells were fixed using 2.5% glutaraldehyde in phosphate-buffered saline (PBS) at pH 7.2, contrasted with 1% osmium tetroxide (Sigma-Aldrich) and 1% carbohydrazide (Sigma-Aldrich). Subsequently, cells were dehydrated in increasing concentrations of hexylene glycol (Sigma-Aldrich). Drying was performed using a Polaron CPD 7501 critical point dryer (Quorum Technologies, Laughton, UK).
4
SEM Analysis of Bacterial Samples
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A scanning electron microscopy (SEM) analysis of bacterial samples was performed by means of an FEI Quanta 200 electron microscope (FEI Co., Hillsboro, OR, USA). The bacteria samples were rinsed in PBS (0.01 M, pH 7.2; P4417, Sigma-Aldrich, Poznań, Poland) and then fixed in 2.5% glutaraldehyde (G5882, Sigma) for 1 h, washed twice with 0.1 M PBS (0.01 M, pH 7.2; P4417, Sigma), and placed on aluminum SEM stubs. The SEM stubs were kept in a moist atmosphere for 1 h, washed with PBS (0.01 M, pH 7.2; P4417, Sigma), postfixed in 1% osmium tetroxide (75632, Sigma) for 1 h, rinsed in distilled water, and dehydrated in graded ethanol. After critical point drying with liquid CO2 in a vacuum apparatus (Polaron CPD 7501, Quorum Technologies, Newhaven, East Sussex, UK) and coating with a 5.12 nm gold layer gold using an EM ACE 600 high-vacuum sputterer (Leica Microsystems, Wetzlar, Ger-many), the samples were inspected by SEM at 5 KeV (Quanta 250 FEG SEM, FEI, Hillsboro, OR, USA).
5
SEM Analysis of Red Blood Cells
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Scanning electron microscopy (SEM) analysis of RBC was performed by means of an FEI Quanta 200 electron microscope (FEI Co., Hillsboro, OR, USA). The blood samples were rinsed in PBS (0.01 M, pH 7.2; P4417, Sigma) then fixed in 2.5% glutaraldehyde (G5882, Sigma) for 1 h, washed twice in 0.1 M PBS (0,01 M, pH 7.2; P4417, Sigma) and placed on aluminium SEM stubs. The SEM stubs were kept in a moist atmosphere for 1 h, washed in PBS (0.01 M, pH 7.2; P4417, Sigma), post-fixed in 1% osmium tetroxide (75632, Sigma) for 1 h, rinsed in distilled water and dehydrated in graded ethanols. After critical point drying with liquid CO2 in a vacuum apparatus (Polaron CPD 7501, Quorum Technologies, Newhaven, East Sussex, UK) and coating with gold-palladium (JEE-4C, JEOL Ltd., Tokyo, Japan), the samples were inspected by SEM at 1 KeV (FEI Quanta 200).
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