For floral trait analyses of V. reichenbachiana, CH and CL flowers were fixed in acetic acid–alcohol (FAA) (40% formaldehyde, glacial acetic acid and 70% ethanol, 0.5/0.5/9, v/v/v; Avantor Performance Materials Poland S.A., Gliwice, Poland) or in 4% glutaraldehyde (Avantor Performance Materials Poland S.A., Gliwice, Poland) in 0.1 M Na cacodylate buffer (Sigma-Aldrich, Saint Louis, MO, USA), dehydrated in increasing concentrations of ethanol and dried at the critical point of carbon dioxide. Samples were glued onto holders, coated in gold (Jeol JFC-1100E ion sputter [Jeol, Tokyo, Japan] or SPIMODULE Sputter Coater [Structure Probe, Inc., Chester, PA, USA], depending on sample) and examined in a Jeol JSM-5410 SEM (Jeol, Tokyo, Japan) or in a Philips XL 30 SEM (Philips Electron Optics B.V., Eindhoven, The Netherlands).
0.1 m na cacodylate buffer
Manufactured by Merck Group
Sourced in Poland, United States, Germany
0.1 M Na-cacodylate buffer is a laboratory solution used as a buffer in various biochemical and biological applications. It maintains a stable pH environment within the range of 6.0 to 7.4, making it suitable for a variety of experiments and analyses. The solution is composed of sodium cacodylate, a sodium salt of cacodylic acid, which acts as the buffering agent.
Automatically generated - may contain errors
Lab products found in correlation
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Evo ma10 scanning electron microscope, by Zeiss (1 mentions)
Glutaraldehyde, by Merck Group (1 mentions)
Ethanol, by Merck Group (1 mentions)
Ultracut uct microtome, by Leica (1 mentions)
Osmium tetroxide, by Serva Electrophoresis (1 mentions)
Auriga crossbeam workstation, by Zeiss (1 mentions)
4 protocols using 0.1 m na cacodylate buffer
1
Floral Trait Analysis of V. reichenbachiana
2
Colitis Induction and Intestinal Analysis
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Mice were sacrificed by cervical dislocation on the seventh day of colitis induction. The intestines were excised and carefully rinsed with saline. A 30-mm section of colon, which was considered to begin at a point 10 mm away from the caecum, was cut out and weighed. The colon was then dissected into two portions, with 10 mm (sample 1) allotted for histological analysis and 5 mm (sample 2) allotted for transmission electron microscopy (TEM). Sample 1 was fixed in 4% paraformaldehyde (Sigma-Aldrich; Merck KGaA) in 4˚C for 24 h, while sample 2 was fixed in 2.5% paraformaldehyde and 2.5% glutaraldehyde in 0.1 M Na-Cacodylate buffer (pH 7.4; all from Sigma-Aldrich; Merck KGaA) in 4˚C for 24 h. In total, 15 control and 15 UC tissue samples were collected to undergo histological analysis and TEM evaluation.
3
SEM Imaging of Bacterial Biofilms on TPU
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Bacteria were incubated on TPU polymeric films for 24 h at 37 °C. The films were washed carefully with phosphate buffer solution (PBS) 1X and fixed with 2.5% (v/v) glutaraldehyde (Sigma-Aldrich, St. Louis, SM, USA) in 0.1 M Na-cacodylate buffer (Sigma-Aldrich, St. Louis, SM, USA) (pH 7.2), for 1 h at 4 °C. After two washes with Na-cacodylate, to remove excess of glutaraldehyde, S. aureus samples were dehydrated using increasing concentrations of ethanol (25, 50, 75%) for 5 min and two washes of 96% ethanol for 10 min. E. coli samples were dehydrated just with two washes of 96% ethanol (Merck Life Science S.r.l., Milano, Italy) for 10 min. The samples were lyophilized for 3 h using a K-850 apparatus (Emitech Ltd, Ashford, UK) and placed on a mounting base. Finally, TPU films were sputter coated with gold and investigated using a Zeiss EVO-MA10 scanning electron microscope (Carl Zeiss, Oberkochen, Germany), 20 kV acceleration voltage.
4
FIB-SEM Serial Sectioning of Larval Tissues
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For focused ion beam scanning electron microscopy (FIB-SEM) serial sectioning, second- and third-instar wild-type Canton S larvae were used. After rinsing in phosphate-buffered saline, the anterior half of the larva was incubated in fixative [2% formaldehyde with 2.5% glutardialdehyde in 0.1 M Na-cacodylate buffer (pH 7.4); Sigma-Aldrich, Germany] for 30 to 90 min. Then, the head region was cut off and incubated in fresh fixative for 90 min. Samples were washed in Na-cacodylate, followed by postfixation in 1% osmium tetroxide (SERVA Electrophoresis GmbH, Germany) for 2 hours at 48°C in the dark. En bloc staining was carried out with 1% uranyl acetate and 1% phosphotungstic acid in 70% ethanol in the dark overnight before continuing the alcohol dehydration the next day. Samples were transferred to propylene oxide before embedded in Spurr (Plano GmbH, Germany) using ascending Spurr concentrations diluted in propylene oxide for optimal tissue infiltration. Polymerization was carried out at 65°C for 72 hours. Blocks were trimmed using an Ultracut UCT microtome (Leica, Germany), mounted on conventional SEM stubs, and sputtered with 80- to 100-nm platinum. FIB-SEM serial sectioning was carried out using a field-emission SEM Auriga CrossBeam workstation (Zeiss, Germany). FIB fine milling was carried out with 500 pA.
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