Patient samples were analyzed following both reduced osmium tetroxide and ruthenium tetroxide postfixation protocols (Elias, 1996 (link); Gruber et al., 2011 (link)). Murine skin biopsies were fixed in 4% glutaraldehyde (TAAB, Berks, UK) in 0.1 M sodium phosphate and incubated with 1% osmium tetroxide (TAAB) 1.5% potassium ferricyanide overnight at 4 °C, then in 1% tannic acid (TAAB) overnight before serial dehydration. Further steps and imaging were as described (Banushi et al., 2016 (link)).
Glutaraldehyde
Manufactured by TAAB
Sourced in United Kingdom, United States
Glutaraldehyde is a chemical compound used in various laboratory applications. It is a colorless, oily liquid with a pungent odor. Glutaraldehyde is commonly employed as a fixative and cross-linking agent in electron microscopy, histology, and tissue preservation processes.
Automatically generated - may contain errors
Lab products found in correlation
Osmium tetroxide, by TAAB (9 mentions)
Paraformaldehyde (pfa), by TAAB (5 mentions)
Ceta 16m ccd camera, by Thermo Fisher Scientific (5 mentions)
Talos l120c, by Thermo Fisher Scientific (4 mentions)
Spurr s resin, by TAAB (4 mentions)
Paraformaldehyde (pfa), by Merck Group (2 mentions)
Pelco biowave pro microwave, by Ted Pella (2 mentions)
Aqueous osmium tetroxide, by TAAB (2 mentions)
Triton x 100, by Merck Group (2 mentions)
Jsm 6301f, by JEOL (2 mentions)
Osmium tetroxide, by Agar Scientific (2 mentions)
Sodium dodecyl sulfate (sds), by Merck Group (2 mentions)
Paraformaldehyde (pfa), by Electron Microscopy Sciences (2 mentions)
Nitrocellulose sheets, by GE Healthcare (2 mentions)
N n methylene bisacrylamide, by Promega (2 mentions)
Electrophoresis equipment, by Hoefer (2 mentions)
Uranyl acetate, by TAAB (2 mentions)
Tecnai g2 spirit, by Thermo Fisher Scientific (2 mentions)
Epon 812, by TAAB (2 mentions)
Jem 1011, by JEOL (2 mentions)
54 protocols using glutaraldehyde
1
Ultrastructural Analysis of Murine Skin
2
Ultrastructural Analysis of Mouse Corpus Callosum
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Mice were intracardially perfused with 4% PFA (w/v) and 2% glutaraldehyde (v/v; TAAB Laboratories) in 0.1 M phosphate buffer. Tissue was post-fixed overnight at 4 °C and transferred to 1% glutaraldehyde (v/v) until embedding. Tissue sections (1 mm) were post-fixed in 1% osmium tetroxide and dehydrated before processing into araldite resin blocks. Next, 1 μm microtome-cut sections were stained with a 1% toluidine blue/2% sodium borate solution before bright-field imaging using a Zeiss Axio microscope. Ultrathin sections (60 nm) were cut from corpus callosum samples, stained with uranyl acetate and lead citrate, and grids imaged on a JEOL transmission electron microscope. Axon diameter, myelin and inner tongue thickness were calculated from a measured area based on assumption of circularity using Fiji/ImageJ (Fiji.sc) (diameter = 2 × √[area/π]), with 100–200 axons per animal analysed.
3
Pine Needle Chloroplast Ultrastructure Analysis
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Thin slices (< 0.5 mm) from the middle region of pine needles were cut in tap water and fixed in 4% paraformaldehyde, 2.5% glutaraldehyde (TAAB Laboratories, Aldermaston, England) in 0.1 M or 0.05 M (May and June) sodium cacodylate buffer, pH 7.4 (TAAB Laboratories, Aldermaston, England). Thoroughly washed samples were post-fixed in 1% osmium tetroxide (TAAB Laboratories, Aldermaston, England). The fixed material was dehydrated in ethanol series with increasing concentrations and propylene oxide and finally embedded in Spurr resin (TAAB Laboratories, Aldermaston, England). Ultrathin sections (70 nm) were post contrasted in uranyl acetate and Reynolds lead citrate and further examined with Talos 120 C electron microscope (FEI, Eindhoven, The Netherlands) operating at 120 kV. Micrographs were acquired with a Ceta 16 M CCD camera (FEI, Eindhoven, The Netherlands) using TEM Image & Analysis software ver. 4.14 (FEI, Eindhoven, The Netherlands). The chloroplast ultrastructure was analyzed from the electron micrographs by measuring the average number of chloroplasts per cell, average number of grana per chloroplasts and average number of appressed thylakoids per grana stack (Ng)65 ,66 (link).
4
Stabilizing FtsZ Polymers for Binding Assays
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FtsZ (10–15
μM) was assembled in HEPES buffer, 10 mM MgCl2, 50
μM GMPCPP at 25 °C for 10 min, and then 0.15% (v/v) glutaraldehyde36 (link) (distilled grade for microscopy, TAAB Laboratories,
U.K.) was added to the solution that was incubated at 25 °C for
10 min more. (This was the minimal glutaraldehyde concentration determined
to stabilize FtsZ polymers specifically binding probe 1.) The remains
of the cross-linking agent were quenched by adding 60 mM NaBH4, the sample was incubated on ice for 10 min and degassed.36 (link) Cross-linked polymers were centrifuged for 10
min at 8200g (5000 rpm) and 4 °C in 15 mL Falcon
tubes employing a Rotina 380R (Hettich) centrifuge, the supernatant
was removed and the pellet was resuspended in the same volume of HEPES
buffer, 10 mM MgCl2, containing 5 μM GMPCPP. Fixed
FtsZ polymers were active in binding assays after more than 2 days
at 4 °C; they could also be frozen in liquid nitrogen with a
small loss of binding capacity. However, they were observed to precipitate
above 20 μM FtsZ concentration. Cross-linked polymers of SaFtsZ
were similarly prepared.
μM) was assembled in HEPES buffer, 10 mM MgCl2, 50
μM GMPCPP at 25 °C for 10 min, and then 0.15% (v/v) glutaraldehyde36 (link) (distilled grade for microscopy, TAAB Laboratories,
U.K.) was added to the solution that was incubated at 25 °C for
10 min more. (This was the minimal glutaraldehyde concentration determined
to stabilize FtsZ polymers specifically binding probe 1.) The remains
of the cross-linking agent were quenched by adding 60 mM NaBH4, the sample was incubated on ice for 10 min and degassed.36 (link) Cross-linked polymers were centrifuged for 10
min at 8200g (5000 rpm) and 4 °C in 15 mL Falcon
tubes employing a Rotina 380R (Hettich) centrifuge, the supernatant
was removed and the pellet was resuspended in the same volume of HEPES
buffer, 10 mM MgCl2, containing 5 μM GMPCPP. Fixed
FtsZ polymers were active in binding assays after more than 2 days
at 4 °C; they could also be frozen in liquid nitrogen with a
small loss of binding capacity. However, they were observed to precipitate
above 20 μM FtsZ concentration. Cross-linked polymers of SaFtsZ
were similarly prepared.
5
Visualizing Recombinant Protein in E. coli
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Induced E. coli cells containing recombinant FilP in pET28a plasmid were harvested and fixed with 2.5% glutaraldehyde (TAAB Laboratories Equipment) and 4% paraformaldehyde (Thermo Fisher Scientific), embedded in LR white resin (Sigma-Aldrich), and processed with an ultramicrotome (Leica EM UC7) into 70-nm thin sections. Sections were contrasted using uranyl acetate (Polysciences) and lead citrate (Thermo Fisher Scientific). Images were collected with Talos L120 TEM (FEI) using Ceta CMOS detector (FEI).
6
Ultrastructural Analysis of Cells
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Cells were seeded onto a 60-mm dish at 1×106 cells/dish and treated with various reagents for 48 h. Then, the cells were fixed for 1 h at 4°C with 2.5% glutaraldehyde (TAAB Laboratories Equipment, Ltd.) in 0.1 M phosphate buffer (pH 7.3) (prepared with monobasic sodium phosphate and dibasic sodium phosphate; Wako Pure Chemical Industries), and fixed subsequently for 1 h at RT in 1% osmium tetroxide (Nisshin EM Co., Ltd.), dehydrated in graded ethanol (30-100%), and embedded in Quetol 812 epoxy resin (Nisshin EM Co., Ltd.) at 60°C for 2-4 days. ultrathin sections (60 nm) were obtained using an Ultracut J micro-tome (Reichert Jung), and the sections were stained with 4% lead nitrate (RT, 5 min) and saturated uranium acetate (RT, 10 min) and imaged using a transmission electron micro-scope JEM-1200EX II (JEOL, Ltd.) (magnification ranging from ×1,000 to ×10,000). All images were captured on films (Electron-microscopic film FG; Fujifilm).
7
Tissue Fixation and Sectioning for Microscopy
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Animals were anaesthetised with either sodium pentobarbital (50 mg/kg, i.p.) or chloral hydrate (350 mg/kg) and transcardially perfused with saline followed by either 2% or 4% paraformaldehyde (wt/vol, PFA, Sigma-Aldrich), 15% saturated picric acid (vol/vol, Sigma-Aldrich) and 0.05% glutaraldehyde (wt/vol, distilled grade, TAAB Laboratories Equipment Ltd) in 0.1 M phosphate buffer (PB) at pH 7.2. After brain removal, coronal sections were produced (70 μm nominal thickness, VT1000s vibratome, Leica Instruments). Sections were stored in 0.1 M PB with 0.05% sodium azide at 4 °C.
8
Epon and Lowicryl Embedding Protocols
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For Epon embedding, cell cultures were fixed for 1 h at 4°C in 2% glutaraldehyde (Taab Laboratory Equipment) in 0.1 M phosphate buffer, pH 7.3. During fixation, cells were scraped off from the plastic substratum and centrifuged at 5,000g for 15 min. Cell pellets were dehydrated in increasing concentrations of ethanol and embedded in Epon. Polymerization was carried out for 48 h at 64°C. Ultrathin sections were collected on Formvar–carbon–coated copper grids (200 mesh) and stained briefly with standard uranyl acetate and lead citrate solutions.
Embedding in Lowicryl K4M (Chemische Werke Lowi) was carried out on Vero cells fixed either in 4% formaldehyde (Merck) or in 2% glutaraldehyde at 4°C. Cell pellets were equilibrated in 30% methanol and deposited in a Leica EM AFS2/FSP automatic reagent handling apparatus (Leica Microsystems). Lowicryl polymerization under UV was for 40 h at −20°C followed by 40 h at +20°C. Ultrathin sections of Lowicryl-embedded material were collected on Formvar–carbon–coated gold grids (200 mesh) and stored until use.
Embedding in Lowicryl K4M (Chemische Werke Lowi) was carried out on Vero cells fixed either in 4% formaldehyde (Merck) or in 2% glutaraldehyde at 4°C. Cell pellets were equilibrated in 30% methanol and deposited in a Leica EM AFS2/FSP automatic reagent handling apparatus (Leica Microsystems). Lowicryl polymerization under UV was for 40 h at −20°C followed by 40 h at +20°C. Ultrathin sections of Lowicryl-embedded material were collected on Formvar–carbon–coated gold grids (200 mesh) and stored until use.
9
Light and Electron Microscopy of Inflorescence Stems
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For light microscopy, the bottom 1 cm of 10‐week‐old inflorescence stems was cut into 70‐μm cross‐sections with a vibratome. The cross‐sections were stained for 1 min in 0.01% toluidine blue and washed twice with water. Images of the cross‐sections were captured using an Axioplan microscope (Carl Zeiss, Oberkochen, Germany) with a camera attachment.
For the TEM, the bottom 1 cm of 10‐week‐old inflorescence stems was cut into small pieces and fixed in 2.5% glutaraldehyde (TAAB Laboratories, Aldermaston, UK) and 4% paraformaldehyde in 0.1m sodium cacodylate buffer, and then further post‐fixed in 1% aqueous osmium tetroxide (TAAB Laboratories). Samples were dehydrated in ethanol and propylene oxide and finally embedded in Spurr's resin (TAAB Laboratories). All of the samples were processed using a Pelco Biowave Pro+ microwave tissue processor (Ted Pella, Redding, CA, USA). The 70‐nm ultrathin sections were post‐contrasted in uranyl acetate and Reynolds' lead citrate and further examined with a Talos L120C (FEI, Eindhoven, The Netherlands) operating at 120 kV. Micrographs were acquired with a Ceta 16M CCD camera (FEI) using tem image & analysis , version 4.17 (FEI).
For the TEM, the bottom 1 cm of 10‐week‐old inflorescence stems was cut into small pieces and fixed in 2.5% glutaraldehyde (TAAB Laboratories, Aldermaston, UK) and 4% paraformaldehyde in 0.1
10
Decellularized Bovine Tissue Matrix Characterization
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Bovine tissue strips were imaged using scanning electron microscopy (SEM) to observe any architectural damage caused to the dCELL tissues’ structural matrix following decellularization compared to untreated cellular BTM (n = 3). Bovine tissue strips were fixed in 2.5% v/v Glutaraldehyde (TAAB Laboratories Equipment Ltd, Reading, UK) for 24 h at 4 °C. The strips were dehydrated in ascending concentrations of Ethanol (50–100% v/v; (Decon Laboratories Inc., King of Prussia, PA, USA)), critical point dried with Hexamethyldisilazane (HMDS; Sigma-Aldrich, Gillingham, UK) and mounted onto carbon-tabbed SEM stubs (Agar Scientific Ltd., Essex, UK). Mounted strips were Gold/Palladium (AuPd) sputter-coated to increase electrical conductivity and imaged using a HITACHI TM4000 Plus tabletop SEM at high vacuum with a 15 kV electron beam.
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