For histological analysis, constructs were fixed with 4% (w/v) paraformaldehyde including 100 mM sodium cacodylate trihydrate (Sigma) and 10 mM CaCl2, and incubated at 4°C in 50 mM BaCl2 solution containing 100 mM sodium cacodylate trihydrate to stabilize the alginate[10 ]. Then the constructs were paraffin-embedded and sectioned at 6 μm. Safranin-O staining was performed to assess glycosaminoglycan production. To confirm that the chondrocytes in this study were not dedifferentiated, we performed immunohistochemistry with type II collagen and type I collagen staining. Antigen was retrieved with 5 mg/mL of hyaluronidase in PBS for 30 min at 37°C. The 6-μm thick sections were incubated with primary antibodies against type II collagen and type I collagen overnight at 4°C (SC-25974, SC-7764, Santa Cruz Technology, Santa Cruz, CA, USA). Thereafter, the sections were treated sequentially with a Texas Red-conjugated secondary antibody (SC-2783, Santa Cruz Technology, Santa Cruz, CA, USA) for 30 min at room temperature, followed by counterstaining with Hoechst 33342 (Pierce, Rockford, IL, USA). Photography was performed with a Nikon E800 microscope (Nikon, Melville, NY, USA).
Sodium cacodylate trihydrate
Manufactured by Merck Group
Sourced in United States, Germany, France
Sodium cacodylate trihydrate is a chemical compound used as a buffer in various laboratory applications. It is a crystalline solid that is soluble in water and commonly used to maintain a specific pH level in buffers and solutions. The core function of sodium cacodylate trihydrate is to stabilize the pH of laboratory samples and reagents.
Automatically generated - may contain errors
Lab products found in correlation
Sodium hydroxide (naoh), by Merck Group (3 mentions)
Glutaraldehyde solution, by Merck Group (3 mentions)
Agar 100, by Agar Scientific (3 mentions)
Nitric acid, by Merck Group (3 mentions)
Paraformaldehyde (pfa), by Merck Group (3 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (2 mentions)
Em 900, by Zeiss (2 mentions)
Sucrose, by Merck Group (2 mentions)
Glutaraldehyde, by Electron Microscopy Sciences (2 mentions)
Osmium tetroxide, by Merck Group (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Iron 2 chloride tetrahydrate, by Merck Group (2 mentions)
Ethanol, by Avantor (2 mentions)
Polyvinylpyrrolidone, by Merck Group (2 mentions)
Diethylene glycol, by Merck Group (2 mentions)
Hydrazine hydrate, by Merck Group (2 mentions)
N methyldiethanolamine, by Merck Group (2 mentions)
Ammonium sulfide solution, by Merck Group (2 mentions)
Iron 3 chloride hexahydrate, by Avantor (2 mentions)
Copper 2 nitrate hemi pentahydrate, by Merck Group (2 mentions)
19 protocols using sodium cacodylate trihydrate
1
Chondrocyte Histological Analysis Protocol
2
Chitosan-based Biomaterials Synthesis and Characterization
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Chitosan
with low/medium/high (CS_L/M/H)
average molecular weight (Mw ∼ 369 ± 4; 1278 ± 8;
2520 ± 9 kDa, respectively) was purchased from Sigma-Aldrich
and used as received. Chitosan L and M were obtained from chitin of
shrimp shells whereas chitosan H was obtained from chitin of crab
shells. The deacetylation degree for CS_L/M/H was 86 ± 3%; 89
± 2%; 85 ± 3%, respectively.54 (link) Aqueous solutions of acetic acid (99.8% Sigma-Aldrich) were used
as the solvent. Gold(III) chloride trihydrate (≥99.9%; 48.5–50.25%
Au), sodium hydroxide (anhydrous, ≥98%), thiazolyl blue tetrazolium
bromide (98%) and the LDH (lactate dehydrogenase) assay kit were also
supplied by Sigma-Aldrich. Dimethyl sulfoxide (DMSO) and methanol
were purchased from Chempur. Phosphate-buffered saline (PBS) without
Ca and Mg was purchased from PAA The Cell Culture Company. Dulbecco’s
modified Eagle’s medium (DMEM) high in glucose (4.5 g/L) withl -glutamine with and without phenol red was used in cell culturing
and was supplied by Thermo Scientific. Materials for bacteria culturing
were purchased from BIOMED (broth) and BIOCORP (agar). Sucrose, sodium
cacodylate trihydrate (approximately 98 wt %), glutaraldehyde solution
(50 wt % in water) and methanol anhydrous 99.8 wt % (Sigma-Aldrich)
were used to fix and dehydrate the cells before scanning electron
microscopy (SEM) visualization.
with low/medium/high (CS_L/M/H)
average molecular weight (Mw ∼ 369 ± 4; 1278 ± 8;
2520 ± 9 kDa, respectively) was purchased from Sigma-Aldrich
and used as received. Chitosan L and M were obtained from chitin of
shrimp shells whereas chitosan H was obtained from chitin of crab
shells. The deacetylation degree for CS_L/M/H was 86 ± 3%; 89
± 2%; 85 ± 3%, respectively.54 (link) Aqueous solutions of acetic acid (99.8% Sigma-Aldrich) were used
as the solvent. Gold(III) chloride trihydrate (≥99.9%; 48.5–50.25%
Au), sodium hydroxide (anhydrous, ≥98%), thiazolyl blue tetrazolium
bromide (98%) and the LDH (lactate dehydrogenase) assay kit were also
supplied by Sigma-Aldrich. Dimethyl sulfoxide (DMSO) and methanol
were purchased from Chempur. Phosphate-buffered saline (PBS) without
Ca and Mg was purchased from PAA The Cell Culture Company. Dulbecco’s
modified Eagle’s medium (DMEM) high in glucose (4.5 g/L) with
and was supplied by Thermo Scientific. Materials for bacteria culturing
were purchased from BIOMED (broth) and BIOCORP (agar). Sucrose, sodium
cacodylate trihydrate (approximately 98 wt %), glutaraldehyde solution
(50 wt % in water) and methanol anhydrous 99.8 wt % (Sigma-Aldrich)
were used to fix and dehydrate the cells before scanning electron
microscopy (SEM) visualization.
3
Tissue Preparation for TEM Imaging
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Tissue samples were fixed with 2.5% glutaraldehyde (Assing Spa, R1012) in 0.1 M cacodylate buffer for 1 h at 4°C (sodium cacodylate trihydrate, Sigma-Aldrich, C4945), and postfixed in 1% osmium tetroxide (Sigma-Aldrich, 75632) in 0.1 M cacodylate buffer for 1 h. Samples were then dehydrated in graded ethanol and embedded in Epon resin (AGAR 100, Agar Scientific R1045). Ultrathin sections were stained with 2% uranyl acetate (Sigma-Aldrich, 73943) and observed under a Zeiss EM900 transmission electron microscope. Images were captured digitally with a Mega View II digital camera (SIS; Zeiss).
4
Nanofiber Scaffold Preparation for SEM Imaging
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Nanofiber scaffolds containing T17b eEPCs were washed with PBS first and fixed in multiple steps, starting with incubation in a 0.2 M sodium cacodylate trihydrate (Sigma Aldrich, Schnelldorf, Germany) buffer, containing 0.1% glutaraldehyde (Carl Roth, Karlsruhe, Germany), 2% paraformaldehyde (Carl Roth), and 5% sucrose (Sigma Aldrich, Schnelldorf, Germany) for 1 h at room temperature. In a second step, samples were placed in a 0.2 M cacodylate buffer, containing 0.3% glutaraldehyde and 3% paraformaldehyde for 1 h, followed by a series of ethanol dilutions in rising concentration, ending at 100 vol.%.
Fixed samples were then critical point dried with EM CPD300 (Leica Microsystems, Wetzlar, Germany), removed from Minusheets, gold-palladium sputtered, and examined using an AURIGA® scanning electron microscope (SEM) (Carl Zeiss, Jena, Germany).
Fixed samples were then critical point dried with EM CPD300 (Leica Microsystems, Wetzlar, Germany), removed from Minusheets, gold-palladium sputtered, and examined using an AURIGA® scanning electron microscope (SEM) (Carl Zeiss, Jena, Germany).
5
Ultrastructural Analysis of Kidney Organoids
6
Ultrastructural Analysis of Aortic Arch
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Aortic arch specimens (1 mm3) were fixed in 4% glutaraldehyde (EMD Chemicals Inc, Gibbstown, NJ, USA) and 0.1 M sodium cacodylate trihydrate (Sigma-Aldrich, Germany). Fixed segments were ultrathin-sectioned and prepared according to standard procedure. Electron microscopy was performed using a FEI Tenmai Biotwin transmission electron microscope.
7
Chloroplast Morphometrics via TEM
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Samples for leaf morphometrics were also used for chloroplast morphometrics. Some 5 mm fragmens were sampled from each plant, and were fixed in Karnovsky buffer77 for 30 days, then washed three times in cacodylate buffer (sodium cacodylate trihydrate, part number C0250, Sigma Aldrich, St. Louis, USA) and post-fixed with 1% (v/v) osmium tetroxide (part number 201030, Sigma Aldrich, St. Louis, USA) in 0.05 M phosphate buffer, dehydrated in a series of increasing acetone concentration, infiltrated and polymerized in low viscosity epoxy resin78 (link). Cross sections of 70 nm thickness were obtained using an ultramicrotome (Power Tome-X, RMC Products, Boeckeler Instruments, Inc., Tucson, USA) and stained with uranyl acetate, then lead citrate79 (link). Chloroplast parameters (see Table 2 ) were obtained with a transmission electron microscope, 50 kV, with a coupled digital camera (EM 109, Carl Zeiss Microscopy Ltd., Jena, Germany).
8
Protein Purification and Characterization
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All chemicals
used throughout
the experiments were purchased from commercial suppliers and used
without further purification. HSA and BSA, purchased from Sigma-Aldrich,
were diluted in Milli-Q water to a final concentration of 0.2 mM.
Stock solutions of Myoglobin,l -glutathione, l -arginine,
lysozyme,l -cysteine, chymotrypsin, and chymotrypsinogen
A were prepared by dissolving the samples in Milli-Q water until the
desired concentration was reached. Sodium cacodylate trihydrate (0.05
M), supplied by Sigma-Aldrich, was used to control the pH of the solutions
(pH 7.2).
used throughout
the experiments were purchased from commercial suppliers and used
without further purification. HSA and BSA, purchased from Sigma-Aldrich,
were diluted in Milli-Q water to a final concentration of 0.2 mM.
Stock solutions of Myoglobin,
lysozyme,
A were prepared by dissolving the samples in Milli-Q water until the
desired concentration was reached. Sodium cacodylate trihydrate (0.05
M), supplied by Sigma-Aldrich, was used to control the pH of the solutions
(pH 7.2).
9
Scanning Electron Microscopy of hDFs
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hDFs were cultured with 4.2 F MCMs on tissue culture-treated Thermanox Plastic Coverslips (Nunc) in DMEM with 10 v/v% FBS and 1000 U/mL P/S for 2 days. Cells were fixed on coverslips with 4% paraformaldehyde for 10 min at RT. Cells were further fixed in 0.7 M sodium cacodylate trihydrate (Sigma Aldrich) with 3 mM magnesium chloride (Sigma Aldrich) and 1.5 v/v% glutaraldehyde (Sigma-Aldrich) in DI water for 2 hrs at RT. Fixed cells were washed in 0.7 M sodium cacodylate with 2.5 w/v% sucrose 2X. Washed cells were dehydrated in 10 min subsequent incubations in 30, 50, 70, 95% ethanol in DI water. Dehydration was completed in 10 min subsequent incubations in 30, 50, 70, 95% hexamethyldisilazne (Sigma-Aldrich) in ethanol. Dehydrated cells were imaged on a LEO 1530 scanning electron microscope (Gemini) at 3kv. Cells and MCMs were pseudo colored in Adobe Photoshop to improve clarity of MCM and cellular regions.
10
Synthesis and Characterization of Metal Nanoparticles
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All reagents were of analytical purity and used without further purification. Iron(II) chloride tetrahydrate (FeCl2·4H2O, 99%), sodium hydroxide (NaOH, 99.99%), diethylene glycol (DEG, 99%), N-methyldiethanolamine (NMDEA, 99%), nitric acid (HNO3, 70%), copper(II) nitrate hemi(pentahydrate) (Cu(NO3)2·2.5H2O ≥99.99%), polyvinylpyrrolidone (PVP, Mw 55 kDa), hydrazine hydrate (55%), ammonium sulfide solution ((NH4)2S, 20%), dihydrorhodamine 123 (DHR123 ≥95%), sodium cacodylatetrihydrate (≥98%), glutaraldehyde solution (25% in H2O), hydrogen peroxide solution (30 wt. % in H2O) and formalin solution (10%) were purchased from Sigma-Aldrich (France). Iron(III) chloride hexahydrate (FeCl3·6H2O, 99%) and ethanol were obtained from VWR (France). Alamar BlueTM cell viability reagent was purchased from Thermo-Fisher.
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