Glutaraldehyde

Manufactured by Merck Group

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Glutaraldehyde is a chemical compound used as a fixative and disinfectant in various laboratory applications. It serves as a cross-linking agent, primarily used to preserve biological samples for analysis.

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Lab products found in correlation

Paraformaldehyde (pfa), by Merck Group (208 mentions) Osmium tetroxide, by Merck Group (158 mentions) Bovine serum albumin (bsa), by Merck Group (107 mentions) Ethanol, by Merck Group (95 mentions) Crystal violet, by Merck Group (66 mentions) Formaldehyde, by Merck Group (64 mentions) Triton x 100, by Merck Group (61 mentions) Sodium hydroxide (naoh), by Merck Group (58 mentions) Dimethyl sulfoxide (dmso), by Merck Group (55 mentions) Phosphate buffered saline (pbs), by Merck Group (54 mentions) Chitosan, by Merck Group (54 mentions) Lead citrate, by Merck Group (51 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (51 mentions) Uranyl acetate, by Merck Group (47 mentions) Acetic acid, by Merck Group (46 mentions) Hexamethyldisilazane, by Merck Group (43 mentions) Poly l lysine (pll), by Merck Group (42 mentions) Acetone, by Merck Group (41 mentions) Hydrochloric acid (hcl), by Merck Group (41 mentions) Sodium cacodylate, by Merck Group (37 mentions)

2 338 protocols using glutaraldehyde

Osmium-Staining Procedure for Cell-Seeded Scaffolds

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The osmium-staining
procedure was performed as previously described.^{32 (link)} Briefly, the cell-seeded scaffolds were immersed in 4%
glutaraldehyde (Sigma) for 30 min at RT on each timepoint and then
washed with PBS three times and stored in PBS at 4 °C until use.
Glutaraldehyde-fixed cell-seeded scaffolds were incubated in 0.1%
osmium tetroxide (TAAB) in deionized water for 30 min before dehydrating
in ethanol (Sigma) and hexamethyldisilazane (HDMS, Sigma). Samples
were dried overnight in the fume cupboard and visualized using a Hitachi
TM4000 SEM (Hitachi) at different magnifications.

Gao Y., Bate T.S, & Callanan A. (2023). A Unification of Nanotopography and Extracellular Matrix in Electrospun Scaffolds for Bioengineered Hepatic Models. ACS Applied Bio Materials, 6(6), 2158-2171.

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FE-SEM Sample Preparation Protocol

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For FE-SEM analysis, each material was rinsed thrice with warm PBS, fixed in 3% glutaraldehyde (Sigma-Aldrich, USA) at 4°C overnight, rinsed thrice with PBS, and postfixed with 3% glutaraldehyde for 2 h at RT. The samples were dehydrated in a graded series of ethanol for 2 h and placed into a desiccator until the analysis.

Valdez-Salas B, & Beltrán-Partida E. (2021). Feasibility of Using H3PO4/H2O2 in the Synthesis of Antimicrobial TiO2 Nanoporous Surfaces. Bioinorganic Chemistry and Applications, 2021, 6209094.

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Ultrastructural Analysis of IEL-NK Cells

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Purified CD3⁻/28-4⁺ IEL-NK cells were fixed using 2.5% Glutaraldehyde (Sigma Aldrich, USA) for 4–6 h. The cells were then submerged in proper animal serum and later sliced into 1 mm³ before another fixation in 2.5% Glutaraldehyde at 4°C for 1–2 h. Next, the cells were washed three times with 0.1 M of sodium Cacodylate Buffer (Sigma Aldrich, USA) for 10 min at 400 × g followed by another fixation with 1% Osmium Tetroxide (Sigma Aldrich, USA) at 4°C for 2 h. The cells were then dehydrated for 10 min each in 35, 50, 75, and 95% acetone percentages and 15 min in 100% acetone. The dehydrated samples were placed in a full resin beam capsule and polymerized in the oven at 60°C for 24–48 h. Later, the samples were cut to 1 μm, stained with Toluidine Blue (Sigma-Aldrich, USA), and then the area of interest was examined using a light microscope. Finally, ultrathin sectioning was performed before the cells were stained with uranyl acetate for 15 min and viewed under the Electron Microscope (TEM) (Hitachi H-7100, Japan).

Abdolmaleki M., Yeap S.K., Tan S.W., Satharasinghe D.A., Bello M.B., Jahromi M.Z., Bejo M.H., Omar A.R, & Ideris A. (2018). Effects of Newcastle Disease Virus Infection on Chicken Intestinal Intraepithelial Natural Killer Cells. Frontiers in Immunology, 9, 1386.

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Evaluating MC3T3-E1 Cell Morphology

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MC3T3-E1 cells were seeded in 24-well plates at the density of 2 × 10⁴ cells/cm² with the Zn-HA leaching liquor as above. After 24 h incubation, the cells were washed twice with PBS, then fixed using 2.5% glutaraldehyde (Sigma, USA) at 4°C for 1 h and rinsed three times with PBS to eliminate residual glutaraldehyde. Subsequently, the cells were dehydrated in a graded series of ethanol (30%, 50%, 70%, 90%, and 100%) for 30 min. The MC3T3-E1 cell morphology was observed under a scanning electron microscope (Inspect F, FEI Ltd., Netherlands).

Li Y., Shi X, & Li W. (2017). Zinc-Containing Hydroxyapatite Enhances Cold-Light-Activated Tooth Bleaching Treatment In Vitro. BioMed Research International, 2017, 6261248.

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Ultrastructural Analysis of Fungal Cultures

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The ultrastructural features of fungal WT and mutant cultures were examined using transmission electron microscopy (TEM). Fresh mycelia were cultured in PDB for 24 h at 50 and 37 °C, respectively to remove any medium, and then were incubated with 2.5% glutaraldehyde (Sigma) in phosphate buffer (pH 7.4) overnight at 4 °C. Conidia were cultured in PDA for 7day at 50 °C and 14 day at 37 °C. The samples were pretreated in 4% paraformaldehyde, then processed with 2.5% glutaraldehyde (Sigma) in phosphate buffer (pH 7.4) overnight at 4 °C and post-fixation in 1% OsO4, the samples were dehydrated in a graded ethanol series, embedded in Spurr resin, and stained with 2% uranyl acetate and Reynold's lead solution. The samples were examined under an H-7650 transmission electron microscope (Hitachi).

Chen Y., Yang X., Zhang L., Wu Q., Li S., Gou J., He J., Zhang K., Li S, & Niu X. (2023). Tryptophan-centered metabolic alterations coincides with lipid-mediated fungal response to cold stress. Heliyon, 9(2), e13066.

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Evaluating Cellular Morphology on Anodized Materials

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To evaluate morphological changes in MC3T3-E1 cells seeded on the anodized materials, FE-SEM analysis was carried out as described by others.25 (link) After culturing for periods of 4 h, 24 h, and 21 days of seeding, cells grown on the samples were rinsed three times with PBS (5 min) and fixed in 2.5% w/v glutaraldehyde (Sigma-Aldrich, USA) buffered with 0.1 M sodium cacodylate (Sigma-Aldrich, USA) at 4 °C overnight, washed three times for 5 min in 0.1 M sodium cacodylate buffer and postfixed with 2.5% glutaraldehyde for 2 h at room temperature (RT). The cells were dehydrated in a graded series of ethanol solutions (25%, 50%, 70%, and 100%) for 15 min at each concentration. Finally, the samples were sputter-coated with gold (10-nm gold layer) for 8 s and observed at 5 kV accelerating voltage. The image J software was used to compute the length and density of filopodia on the NTs under the different biochemical serum conditions.37 (link)

Valdez-Salas B., Castillo-Uribe S., Beltran-Partida E., Curiel-Alvarez M., Perez-Landeros O., Guerra-Balcazar M., Cheng N., Gonzalez-Mendoza D, & Flores-Peñaloza O. (2022). Recovering Osteoblast Functionality on TiO2 Nanotube Surfaces Under Diabetic Conditions. International Journal of Nanomedicine, 17, 5469-5488.

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Electrospun Gelatin-Fibrinogen Scaffold Creation

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Gelatin-Fibrinogen flat sheet scaffolds were created by electrospinning. Gelatin extracted from porcine skin and fraction I bovine fibrinogen (Sigma-Aldrich, USA) were mixed at three different percentages: 100% gelatin (100 G), 80% gelatin-20% fibrinogen (80:20 G:F) and 50% gelatin-50% fibrinogen (50:50 G:F) [16 (link)]. The polymeric blends were dissolved in 1,1,1,3,3,3-Hexafluoro-2-propanol (HFP) (Sigma-Aldrich, USA) to create a 10% (w/v) solution under constant stirring. The solutions were loaded into a 5 ml BD syringe with a 23 gauge stainless steel dispensing blunt tip needle (CML supply, USA) attached. The syringe was then loaded onto a NE-1000 single syringe pump (New era pump systems inc., USA) set to a pumping rate of 100 μl/min. The distance from the needle tip to the target was 8 cm. The polymeric solutions were electrospun at a high voltage of 15 kV, onto glass coverslips attached to a metallic target to create fine fibers. The resultant flat sheets were crosslinked in 25% glutaraldehyde (Sigma- Aldrich, USA) in vapor phase for 24 h. The glutaraldehyde was then removed in a convection oven for 24h at 42°C. Additionally, membranes were rinsed with deionized water to remove any crosslinker residues and uncrosslinked gelatin.

Ardila D.C., Tamimi E., Danford F.L., Haskett D.G., Kellar R.S., Doetschman T, & Vande Geest J.P. (2014). TGFβ2 Differentially Modulates Smooth Muscle Cell Proliferation and Migration in Electrospun Gelatin-Fibrinogen constructs. Biomaterials, 37, 164-173.

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Hydrophilic Surface Modification of Microcapillary Film

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Hydrophilic coating of microcapillary film MCF was produced by Lamina Dielectrics Ltd (Billingshurst, West Sussex, UK) from Teflon® FEP (Dupont, USA) using a melt-extrusion process, 29 and consisted of an array of 10 parallel microcapillaries with a mean hydraulic diameter of 206 ± 12.6 μm. The fluoropolymer MCF was subsequently modified by coating the inner surface of the microcapillaries with a permanent hydrophilic layer of PVOH. This involved recirculating at a flow rate of 50 mL h -1 overnight a volume of 100 mL of a 5 mg mL -1 solution of poly(vinyl alcohol) (PVOH) in water (MW 13 000-23 000, >98% hydrolysed for ABO blood grouping experiments; MW 146 000-186 000, >99% hydrolysed for bacteria and MIC testingall from Sigma-Aldrich, UK). A 6 m long fluoropolymer MCF was attached to a FPLC P-500 Pharmacia Biotech pump using Upchurch flangeless tube fittings (Kinesis, UK). The PVOH coating was then crosslinked with glutaraldehyde by manually filling the MCF with a freshly prepared 5 mg mL -1 of PVOH solution containing 5 mM of glutaraldehyde (Sigma-Aldrich, UK) and 5 mM HCl (Sigma-Aldrich, UK) for 2 hours at 37 °C, followed by manual washing with water and drying with multiple changes of air using a 50 mL syringe.

Reis N.M., Pivetal J., Loo-Zazueta A.L., Barros J.M, & Edwards A.D. (2016). Lab on a stick: multi-analyte cellular assays in a microfluidic dipstick. Lab on a chip, 16(15).

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Graphene Nanogrid Immunoassay for Hepatitis B

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For immobilization of anti Hep-B monoclonal antibody, (procured from Sigma Aldrich, St. Louis, MO, USA) the graphene nanogrid structure has been treated with 25%, 10% and 2.5% glutaraldehyde aqueous solutions (procured from Sigma Aldrich) for 2 h, 4 h and 24 h. After treatment, each sample has been rinsed with de-ionized water and for every parameter, optical density (O.D.) measurement has been carried out to estimate the covalent binding of graphene with glutaraldehyde. Then the graphene structure has been incubated with anti Hep-B antibody for 1 h followed by washing with phosphate buffer saline (PBS). This process activates the negatively charged carboxylic groups of graphene and improves the covalent binding of antibodies. After antibody immobilization the nanogrid structure has been incubated with different concentration of Hep-B solution for 10 min. Hep-B solution has been prepared in the range of 50 aM to 10 pM by standard process of serial dilutions [28 (link)]. The binding of antigen on graphene has been schematically represented in Figure 1.

Basu J, & RoyChaudhuri C. (2016). Graphene Nanogrids FET Immunosensor: Signal to Noise Ratio Enhancement. Sensors (Basel, Switzerland), 16(10), 1481.

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Viral Particle Imaging via Electron Microscopy

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Electron microscopy grid preparation was carried out in the BSL4 laboratory. Viral supernatant (2 μL) was deposited directly onto carbon-formvar-coated 300-mesh grids (Polysciences, Warrington, PA, USA) and allowed to dry. The grids were fixed with 2% glutaraldehyde solution for 20 min (unless otherwise stated) and then rinsed by inversion on three sequential drops of ultrapure water. The fixative solution was 2% cacodylate-buffered glutaraldehyde solution prepared using a 0.4 M cacodylate buffer stock combined with 25% glutaraldehyde (Sigma-Aldrich, St. Louis, MO, USA) and ultrapure water.

Alfson K.J, & Griffiths A. (2018). Development and Testing of a Method for Validating Chemical Inactivation of Ebola Virus. Viruses, 10(3), 126.

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