G5882

Manufactured by Merck Group

Sourced in United States, Germany, United Kingdom, Italy

The G5882 is a laboratory equipment product offered by the Merck Group. It is a high-precision instrument designed for use in scientific research and analysis. The core function of the G5882 is to perform measurement and detection tasks, providing accurate and reliable data for a variety of applications.

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G5882-50mL (2 citations)

95 protocols using g5882

Scaffold Fixation and SEM Analysis

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Scaffolds previously used on the cytocompatibility studies were further removed from the plates and fixated for SEM analysis, based on the Utah State University Biological Sample Fixation protocol. In brief, scaffolds were washed with 0.1 M HEPES (Merck^®, PHG0001) 3 times and further fixated in a 2% glutaraldehyde (Merck^®, G5882) buffered solution overnight. The fixative was then removed, and scaffolds were washed with HEPES 3 times, 5 min each, under gentle agitation. From this point, samples were subjected to a crescent series of ethanol (50%, 70%, 95% and 99%) for dehydration, 2–3 times for 10 to 15 min each. Following this, scaffolds were soaked in a crescent series of hexamethyldisilazane (HMDS—Alfa Aesar, A15139)-alcohol solution (1:2; 1:1; 2:1) until complete impregnation in a 98% HMDS solution, 3 times for 15 min. Finally, HMDS was removed from the wells and left to evaporate on an air flow chamber overnight. Samples were coated with Au/Pd by sputtering (SPI Module Sputter Coater) and the SEM/EDX exam was performed using a high resolution (Schottky) Environmental Scanning Electron Microscope with X-ray microanalysis and the Electron Backscattered Diffraction analysis was performed in a Quanta 400 FEG ESEM/EDAX Genesis X4M in high vacuum mode.

Biscaia S., Branquinho M.V., Alvites R.D., Fonseca R., Sousa A.C., Pedrosa S.S., Caseiro A.R., Guedes F., Patrício T., Viana T., Mateus A., Maurício A.C, & Alves N. (2022). 3D Printed Poly( -caprolactone)/Hydroxyapatite Scaffolds for Bone Tissue Engineering: A Comparative Study on a Composite Preparation by Melt Blending or Solvent Casting Techniques and the Influence of Bioceramic Content on Scaffold Properties. International Journal of Molecular Sciences, 23(4), 2318.

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Visualizing Phytoplasma and Endophytes

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A total of four of each fruit, seedling hypocotyl tissue, and two leaf samples tested PCR positive for phytoplasma selected for scanning electron microscopy assessment. The selected plant samples, which were hand sliced to 0.5 to 1 mm thickness, were fixed in 2.5% glutaraldehyde solution (G5882; Merck, Darmstadt, Germany. The fixed samples were dehydrated using graded alcohol concentrations of 20%, 30%, 40%, 50%, 60%, 70%, 80%, and 90%, one time and twice in 100% for 10 min each [31 (link)]. The dehydrated samples were coated with gold particles using a sputter coater (model, SC7620; Quorum Technologies, Lewes, UK). The size and shape of phytoplasma and other endophytic microbial cells were visualized and determined by a scanning electron microscope (Carl Zeiss, EVO 18, Version 6.02).

Kirdat K., Tiwarekar B., Swetha P., Padma S., Thorat V., Manjula K.N., Kavya N., Sundararaj R, & Yadav A. (2022). Nested Real-Time PCR Assessment of Vertical Transmission of Sandalwood Spike Phytoplasma (‘Ca. Phytoplasma asteris’). Biology, 11(10), 1494.

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Senescence Detection via Dimri's Staining

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Dimri’s staining protocol, was used to detect cellular senescence [83 (link)]. Briefly, cells were seeded in a 3.5 cm dish and cultured for at least 72 h. They were washed in Dulbecco’s phosphate-buffered saline (PBS; Sigma-Aldrich, St. Louis, MO, USA, D8537) for 5 min. To fix the cells, a fixation solution containing 0.2% glutaraldehyde solution (Sigma-Aldrich, G5882) and 2% formaldehyde solution (Merck KGaA, Darmstadt, Germany, 104003) was prepared in PBS. After 5 min of fixation, the cells were washed twice with PBS for 5min. Subsequently, the cells were incubated overnight at 37 °C (without CO₂) in the SA-β-Gal staining solution, which contained 5 mM potassium ferricyanide (III) (KGaA, 104973), 5 mM potassium ferrocyanide (II) (Sigma-Aldrich, St. Louis, MO, USA, P9387), 2 mM MgCl₂ (Sigma-Aldrich, M1028), 150 mM NaCl (Sigma-Aldrich, St. Louis, MO, USA, M1028), 0.5 mg/mL 5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside (X-gal; Roche, Basel, Germany, 3117073001), and 40 mM citrate/sodium phosphate buffer (Sigma-Aldrich, St. Louis, MO, USA, S5136), pH = 6. For the analysis, 1000 cells were counted from each sample with a Axiovert 40 CFL bright field microscope. The senescence test was performed at each passage and before the experiments as well as after seeding in some experiments to ensure that the cultures exhibited the same percentage of senescent cells to allow comparison.

Vehns E., Arnold R, & Djabali K. (2022). Impact of MnTBAP and Baricitinib Treatment on Hutchinson–Gilford Progeria Fibroblasts. Pharmaceuticals, 15(8), 945.

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Visualizing Border Cell Migration in Drosophila

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BCs were identified either by the slbo-Gal4-driven GFP expression or by immunostaining with anti-Fas III antibodies (that reveal polar but not outer BCs [6 (link)]) or by X-gal (5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside; #A1007.0001; BioChemica, AppliChem, Germany) staining of slbo¹ (slbo-LacZ) mutants. The latter procedure was performed as follows. Drosophila ovaries were dissected in 1×PBS (1.7 mM KH₂PO₄, 5.2 mM Na₂HPO₄, 150 mM NaCl; pH 7.4) and then fixed in 0.75% glutaraldehyde (#G5882; Merck, Darmstadt, Germany) in 100 mM sodium cacodylate buffer (pH 7.0) (#A2140; PanReac AppliChem, Chicago, IL, USA) for 20 min at room temperature. Next, the ovaries were incubated in a staining solution (10 mM sodium phosphate (pH 7.2), 3.1 mM K₄[Fe(CN)₆], 3.1 mM K₃[Fe(CN)₆], 150 mM NaCl, 1.0 mM MgCl₂, 0.2% X-gal, 0.3% Triton X-100) for 1 h at 37 °C. The migration and completion indexes characterizing BC migration process were calculated as described previously [30 (link)].

Ogienko A.A., Yarinich L.A., Fedorova E.V., Dorogova N.V., Bayborodin S.I., Baricheva E.M, & Pindyurin A.V. (2020). GAGA Regulates Border Cell Migration in Drosophila. International Journal of Molecular Sciences, 21(20), 7468.

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SEM Analysis of Bioceramic Scaffolds

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Further, seeded scaffolds were fixated for SEM analysis, as described in previous works [31 (link),33 (link)]. Scaffolds were rinsed three times with a 0.1M HEPES (Merck^®, PHG0001) buffer solution and left overnight in a fixative solution containing 2% glutaraldehyde (Merck^®, G5882). A dehydration crescent alcohol series (50%, 70%, 95% and 99%) was conducted previously to the incorporation of hexamethyldisilazane (HMDS, Alfa Aesar, A15139). Samples were left overnight to evaporate remaining residues of the reagents.
Following, samples were coated with Au/Pd using sputtering (SPI Module Sputter Coater) for SEM analysis with a high resolution (Schottky) environmental scanning electron microscope with x-ray microanalysis and electron backscattered diffraction analysis, Quanta 400 FEG ESEM/EDAX Genesis X4M, in high vacuum mode.

Guedes F., Branquinho M.V., Biscaia S., Alvites R.D., Sousa A.C., Lopes B., Sousa P., Rêma A., Amorim I., Faria F., Patrício T.M., Alves N., Bugalho A, & Maurício A.C. (2022). Gamma Irradiation Processing on 3D PCL Devices—A Preliminary Biocompatibility Assessment. International Journal of Molecular Sciences, 23(24), 15916.

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Gelatin-coated Coverslips for Cell Culture

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Oregon Green^TM 488–conjugated gelatin-coated (G13186, Thermo Fisher Scientific) coverslips were prepared as previously described⁸¹. In short, coverslips (12 mm diameter, No. 1 thickness; 631-0666, VWR International) were precleaned in 4% nitric acid for 30 min. After washing, the coverslips were coated with 50 µg ml⁻¹ poly-L-lysine (P7890, Merck/Sigma) for 30 min, washed in PBS, and fixed with cold 0.5% glutaraldehyde (G5882, Merck/Sigma) in PBS for 15 min on ice. Subsequently, the coverslips were washed in PBS and coated for 10 min with preheated (44 °C) 10 mg ml⁻¹ unlabelled or Oregon Green^TM 488–conjugated gelatin/2% sucrose in PBS. After coating, the coverslips were washed with PBS and incubated in 5 mg ml⁻¹ sodium borohydride (452882, Merck/Sigma) for 15 min. The coverslips were then washed with PBS, sterilized with 70% ethanol, and equilibrated in serum-containing medium for 1 h at 37 °C before addition of cells.

Wenzel E.M., Pedersen N.M., Elfmark L.A., Wang L., Kjos I., Stang E., Malerød L., Brech A., Stenmark H, & Raiborg C. (2024). Intercellular transfer of cancer cell invasiveness via endosome-mediated protease shedding. Nature Communications, 15, 1277.

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Electron Microscopy Sample Preparation

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Samples were fixed in 2% formaldehyde, 2.5% glutaraldehyde (Sigma Aldrich, G5882) in 0.1 M cacodylate (Sigma Aldrich, C0250) for 24 h. Samples were rinsed with 0.1 M cacodylate, dehydrated in a graded ethanol series, and dried in hexamethyldisilazane (Sigma Aldrich, 440191). Samples were sputter-coated with a thin gold layer (SC7620 Mini Sputter Coater, Quorum Technologies) prior to observation on a scanning electron microscope (JSM-IT200, JEOL).

Dohmen R.G., Hubalek S., Melke J., Messmer T., Cantoni F., Mei A., Hueber R., Mitic R., Remmers D., Moutsatsou P., Post M.J., Jackisch L, & Flack J.E. (2022). Muscle-derived fibro-adipogenic progenitor cells for production of cultured bovine adipose tissue. NPJ Science of Food, 6, 6.

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Muscle Histomorphometry and Fibrosis Analysis

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All animals were sacrificed by a deadly ip injection of ketamine and xylazine and the tibial anterior muscles removed, cleansed of contiguous soft and fat tissue, cut in half in the mid-belly region, and fixated in a solution containing 4% paraformaldehyde (P6148; Sigma-Aldrich, St. Louis, MO, USA), 2.5% sucrose (S0389; Sigma-Aldrich, St. Louis, MO, USA), and 0.1% glutaraldehyde (G5882; Sigma-Aldrich, St. Louis, MO, USA) in PBS (pH 7.2) at 4 °C for 24 h. After fixation, both halves were dehydrated through graded-ethanol solutions, cleared in xylene, and embedded in paraffin blocks. Transverse 5 μm thick sections from each muscle part were cut on a Leica 2125 rotary microtome (Leica Microsystems, Wetzlar, Germany). The created slides were then used for muscle histomorphometry, evaluation of fibrotic-tissue accretion, and immunohistochemistry analysis.

Teixeira E., Garcia J., Bovolini A., Carvalho A., Pacheco J, & Duarte J.A. (2022). Sedentary Behaviour Impairs Skeletal Muscle Repair Modulating the Inflammatory Response. Journal of Functional Morphology and Kinesiology, 7(4), 76.

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Histological Analysis of Vagus Nerve Implants

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Animals were euthanized with an overdose of sodium pentobarbital (400 mg/kg, IP). A 3–6 mm section of vagus nerve was extracted centered on the implanted section. After extraction, the cervical vagus nerves with implanted MINAs were cut (control nerves were left uncut) into three portions, proximal to the implanted area, the implanted area, and distal to the implanted area, and placed in 3% glutaraldehyde (G5882, Sigma) in deionized (DI) water for 24 hours at 4 °C. Samples were then placed in 0.15 M cacodylic acid (Acros, 214971000) in DI water and stored at 4 °C until ready for staining. All subsequent steps occurred at room temperature unless otherwise noted. To begin the staining process, samples were washed three times in 1x PBS (BP3994, Fisher) with each wash lasting 5 minutes. Next, samples were covered with 2% osmium tetroxide (19152, Electron Microscopy Science) for 2 hours. Samples were then triple washed in DI water, followed by triple washing in 1x PBS with each wash lasting 10 minutes. Finally, samples were quadruple washed in 30%, then 50%, and lastly 70% ethanol in DI water for 12 total washes, with each wash taking 5 minutes. The nerve sample containing the implant was then stored in 1x PBS at 4 °C until micro-CT imaging. Proximal, distal, and control nerve samples were stored in 70% ethanol at 4 °C until paraffin processing.

Yan D., Jiman A.A., Bottorff E.C., Patel P.R., Meli D., Welle E.J., Ratze D.C., Havton L.A., Chestek C.A., Kemp S.W., Bruns T.M., Yoon E, & Seymour J. (2022). Ultra-flexible and Stretchable Intrafascicular Peripheral Nerve Recording Device with Axon-dimension, Cuff-less Microneedle Electrode Array. Small (Weinheim an der Bergstrasse, Germany), 18(21), e2200311.

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TEM Assay for Zebrafish Larvae

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The transmission electron microscopy assay was conducted by the molecular medicine testing center of Chongqing Medical University. We prepared samples according to their instructions. Briefly, 3 dpf zebrafish larvae were fixed with 4% glutaraldehyde solution (25% glutaraldehyde in PBS) (G5882, Sigma-Aldrich, USA) for further experiments and photographs.

Zou D., Qin J., Hu W., Wei Z., Zhan Y., He Y., Zhao C, & Li L. (2022). Macrophages Rapidly Seal off the Punctured Zebrafish Larval Brain through a Vital Honeycomb Network Structure. International Journal of Molecular Sciences, 23(18), 10551.

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