1-Anilino-8-naphthalene sulfonic acid (ANS), thioflavin T (ThT), bovine pancreatic insulin, bovine liver catalase, chicken egg white lysozyme, α-glucosidase (α-Gls), dithiothreitol (DTT), isopropyl β-D-1-thiogalactopyranoside (IPTG) and kanamycin were purchased from Sigma. The TEM grid was made of copper and provided by Agar Scientific. β-mercaptoethanol (β-ME), ethylenediaminetetraacetic acid (EDTA) and other chemicals were provided by the Merck Company.
Tem grid
Manufactured by Agar Scientific
Sourced in United Kingdom
TEM grids are used as specimen holders in transmission electron microscopy (TEM) for the analysis of thin samples. They provide a support structure to hold and position the sample during imaging and analysis in the TEM.
Automatically generated - may contain errors
Lab products found in correlation
Copper palladium, by Agar Scientific (2 mentions)
Epon resin lx 112, by Ladd Research Industries (2 mentions)
Osmium tetroxide, by Merck Group (2 mentions)
Chicken egg white lysozyme, by Merck Group (1 mentions)
Bovine pancreatic insulin, by Merck Group (1 mentions)
Kanamycin, by Merck Group (1 mentions)
Bovine liver catalase, by Merck Group (1 mentions)
Isopropyl β d 1 thiogalactopyranoside (iptg), by Merck Group (1 mentions)
Oleic acid, by Thermo Fisher Scientific (1 mentions)
Yb2o3, by Merck Group (1 mentions)
Er2o3, by Merck Group (1 mentions)
Nobf4, by Merck Group (1 mentions)
1 octadecene, by Thermo Fisher Scientific (1 mentions)
Sodium hydroxide (naoh), by Merck Group (1 mentions)
P4707, by Merck Group (1 mentions)
M7145, by Merck Group (1 mentions)
G1393, by Merck Group (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Tecnai t12 biotwin tem, by Thermo Fisher Scientific (1 mentions)
Glutaraldehyde, by Agar Scientific (1 mentions)
16 protocols using tem grid
1
Protein Interaction Assay Protocols
2
Phage Visualization via Transmission Electron Microscopy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Briefly, a small drop of phage suspension containing ∼1 × 107 PFU/mL was applied to a formvar/carbon coated copper transmission electron microscopy (TEM) grid (Agar Scientific, Stansted, United Kingdom) and left for 1 min. Excess liquid was removed with filter paper. A small drop of 2% uranyl acetate (BDH 10288) was applied to the grid surface and left for a further 1 min after which it was removed with filter paper. Grids were left to thoroughly dry before viewing and imaging using a Talos F200c TEM with Gatan Oneview digital camera.
3
Synthesis and Characterization of Upconversion Nanoparticles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Gd2O2S:Yb3+,Er3+ UCNPs (PTIR545) were kindly donated to us by Phosphor Technology Ltd. (UK). For the synthesis of NaYF4:Yb3+,Er3+ UCNPs, Y2O3, Yb2O3, Er2O3 and NaOH were all purchased from Sigma Aldrich UK. Oleic acid, 1-octadecene and NH4F were acquired from Fisher Scientific UK. For the post-synthesis poly(acrylic acid) (PAA)-capping of the UCNPs, both NOBF4 and PAA were purchased from Sigma Aldrich UK. Ferrozine sulphate hydrate, Fe2+ sulphate and MES were purchased from Sigma Aldrich UK, and all salts of competing metal ions were obtained from either Sigma Aldrich UK or Fisher Scientific UK. For the turbid solution measurements, milk powder was purchased from Sainsbury's (UK). The TEM grids used for characterisation were purchased from Agar Scientific (UK).
4
Negative Staining for Nanoparticle Imaging
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Copper/palladium transmission
electron microscopy (TEM) grids (Agar Scientific, UK) were coated
in-house to yield a thin film of amorphous carbon and were subjected
to a glow discharge for 20 s. An aqueous droplet of a copolymer dispersion
(7.0 μL, 0.1% w/w) was placed on freshly treated grids for 1
min and then carefully blotted with filter paper to remove excess
solution. An aqueous droplet of uranyl formate solution (5.0 μL,
0.75% w/w) was placed on each sample-loaded grid for 1 min and then
blotted with filter paper to remove excess stain. This negative staining
protocol was required to ensure sufficient electron contrast. Each
grid was then carefully dried using a vacuum hose. Imaging was performed
at 80 kV using an FEI Tecnai Spirit 2 microscope fitted with an Orius
SC1000B camera. Mean nanoparticle diameters were estimated by digital
image analysis using ImageJ software.
electron microscopy (TEM) grids (Agar Scientific, UK) were coated
in-house to yield a thin film of amorphous carbon and were subjected
to a glow discharge for 20 s. An aqueous droplet of a copolymer dispersion
(7.0 μL, 0.1% w/w) was placed on freshly treated grids for 1
min and then carefully blotted with filter paper to remove excess
solution. An aqueous droplet of uranyl formate solution (5.0 μL,
0.75% w/w) was placed on each sample-loaded grid for 1 min and then
blotted with filter paper to remove excess stain. This negative staining
protocol was required to ensure sufficient electron contrast. Each
grid was then carefully dried using a vacuum hose. Imaging was performed
at 80 kV using an FEI Tecnai Spirit 2 microscope fitted with an Orius
SC1000B camera. Mean nanoparticle diameters were estimated by digital
image analysis using ImageJ software.
5
Staining Nanoparticles on TEM Grids
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Copper/palladium transmission
electron microscopy (TEM) grids (Agar Scientific, UK) were coated
in-house with a thin film of amorphous carbon. Grids were then subjected
to a glow discharge for 30 s to create a hydrophilic surface. Each
0.1%
w/w aqueous diblock copolymer dispersion was deposited as a 5.0 μL
droplet onto a freshly treated grid for 1 min and then blotted with
filter paper to remove excess solution. To stain the deposited nanoparticles,
uranyl formate (5.0 μL of a 0.75% w/w aqueous solution) was
placed on the sample-loaded grid for 20 s and then carefully blotted
to remove excess stain. Each grid was then dried using a vacuum hose.
Imaging was performed using a FEI Tecnai Spirit 2 instrument operating
at 80 kV and equipped with an Orius SC1000B camera.
electron microscopy (TEM) grids (Agar Scientific, UK) were coated
in-house with a thin film of amorphous carbon. Grids were then subjected
to a glow discharge for 30 s to create a hydrophilic surface. Each
0.1%
w/w aqueous diblock copolymer dispersion was deposited as a 5.0 μL
droplet onto a freshly treated grid for 1 min and then blotted with
filter paper to remove excess solution. To stain the deposited nanoparticles,
uranyl formate (5.0 μL of a 0.75% w/w aqueous solution) was
placed on the sample-loaded grid for 20 s and then carefully blotted
to remove excess stain. Each grid was then dried using a vacuum hose.
Imaging was performed using a FEI Tecnai Spirit 2 instrument operating
at 80 kV and equipped with an Orius SC1000B camera.
6
Culturing Murine Brain Endothelial Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The murine brain endothelial cell line bEnd.5 was obtained from the European Collection of Authenticated Cell Cultures (96091930, ECACC, UK). bEnd.5 were grown in high-glucose (4.5 g/L) DMEM (with glutamine and pyruvate) (11885084, Life Technologies, UK) containing 10% FCS (10500064, Life Technologies, UK), 1% penicillin-streptomycin (15070063, Life Technologies, UK), 1× non-essential amino acids (M7145, Merck, Germany), and 5 μM of β-mercaptoethanol (31350010, Life Technologies, UK) in cell culture flasks coated with 2 μg/cm2 of poly-l-lysine (P4707, Merck, Germany) and maintained in an incubator at 37 °C and 5% CO2.
Twenty-four hours before fenestrae induction, bEnd.5 were seeded on glass coverslips or transmission electron microscopy (TEM) grids (Agar Scientific, England, UK) in complete cell culture media. Glass coverslips were coated with 1% bovine gelatin (G1393, Merck, Germany) in 24-well plates. bEnd.5 were seeded at a density of 3.3 × 104 cells per well. Formvar-coated nickel TEM grids were coated with 1% bovine gelatin before seeding 2.5 × 104 cells per grid. Cells were left overnight to facilitate attachment.
Twenty-four hours before fenestrae induction, bEnd.5 were seeded on glass coverslips or transmission electron microscopy (TEM) grids (Agar Scientific, England, UK) in complete cell culture media. Glass coverslips were coated with 1% bovine gelatin (G1393, Merck, Germany) in 24-well plates. bEnd.5 were seeded at a density of 3.3 × 104 cells per well. Formvar-coated nickel TEM grids were coated with 1% bovine gelatin before seeding 2.5 × 104 cells per grid. Cells were left overnight to facilitate attachment.
7
TEM Imaging of Peptide Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Peptides were prepared at 800 µM in PBS with 5% v/v DMSO, and any excessively large aggregates or dust removed by centrifugation (2000 × g, 5 min). Further dilutions (80 µM) were prepared in PBS with 5% v/v DMSO, and the latter diluent was also used to prepare control (peptide-free) grids. Carbon film 200 mesh copper transmission electron microscope (TEM) grids (Agar Scientific Ltd. Stanstead, UK) were initially functionalized using a glow discharger (K100X; Quorum technologies Ltd. Laughton, UK) under a current of 25 mA for 30 sec. These were then placed onto 10 µl droplets of each test sample solution for 1 min, and then transferred onto 10 µl droplets of 1% w/v uranyl acetate for a further minute. Excess solution was removed using filter paper and sample grids were air dried for at least 1 h. Grids were imaged using a Tecnai™ T12 BioTwin TEM (FEI Company. Hillsboro, USA). Images were cropped and scale bars were added using ImageJ. Images are representative of single independent experiments.
8
Optimized Cell Fixation and TEM Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were harvested and fixed for 24 h at 4°C in 3% pentanediol. Cells were washed with phosphate buffer and subsequently postfixed in 1% (v/w) osmium tetroxide (Merck). Samples were dehydrated by successive passages in increasing concentrated ethanol baths (30, 50, 70, 85, and 100%). After embedding in epon resin LX 112 (Ladd Research Industries), ultrathin sections of cell-covered filters were prepared using an 8800 ultrotome III (LKB). TEM analysis used TEM grids (Agar Scientific) covered with nonporous formvar.
9
Transmission Electron Microscopy of Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For transmission electron microscopy (TEM), subconfluent cells were fixed for 2 h at 4 °C in 2.5% (w/v) glutaraldehyde (Agar Scientific) in 0.1 M cacodylate buffer (pH 7.4). Cells were washed with cacodylate buffer and subsequently post-fixed in 1% (v/w) osmium tetroxide (Merck). Samples were dehydrated by successive passages in increasing concentrated ethanol baths (30, 50, 70, 85, and 100%). After embedding in epon resin LX 112 (Ladd Research Industries), ultra-thin sections of cell-covered filters were prepared using an 8800 ultrotome III (LKB). TEM analysis was then performed at 80 keV (FEI Tecnai 10, Philips) using TEM grids (Agar Scientific) covered with non-porous formvar.
10
Embedding Metal Nanoparticles for STXM Examination
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Copper(II) oxide (CuO) nanopowder (<50 nm particle diameter) and Fe0 nanoparticles (25 to 45 nm diameter; both from Sigma-Aldrich) were embedded in an STXM-compatible resin composed of an equimolar mixture of trimethylolpropane triglycidyl ether, 4,4′-methylenebis (2-methylcyclohexylamine), as used during the embedding of the amyloid plaque cores. Semi-thin sections (100 to 500 nm thickness) of the embedded metal standards were cut with a Reichert-Jung Ultra-cut microtome using a diamond blade and were mounted onto TEM grids (Agar Scientific; 100 mesh) for STXM examination. Both the embedding and sectioning of the metal nanoparticles were performed under ambient conditions. These standards were prepared to assess the effect of the embedding process and to confirm that the STXM measurements did not alter the oxidation state of metals embedded within an organic matrix. An additional ferric (Fe3+) iron standard was also prepared in an organic polymer matrix [as described by Brooks et al. (68 )] to further assess the effect on iron oxidation state of successive x-ray beam doses. To avoid cross-contamination of samples, the preparation of metal nanoparticle standards was performed after the preparation and storage of amyloid plaque core materials.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!