In vitro cultured E. multilocularis metacestodes were harvested after 6 months, washed in 100 mM sodium phosphate buffer (pH 7.2) and fixed in phosphate buffer containing 3% paraformaldehyde and 0.05% glutaraldehyde for 2 hours at room temperature. They were then washed in PBS, and sequentially dehydrated in ethanol (30, 50, 70, 90 and 100%) and embedded in LR White acrylic resin (Sigma Aldrich) at -20°C, with three changes of resin during 3 days. Polymerization was done at 60°C overnight. Ultrathin sections were cut by using an ultramicrotome (Reichert and Jung, Vienna, Austria) and were placed on 300 mesh formvar-carbon-coated nickel grids (Plano GmbH, Marburg, Germany). After incubating sections on a drop of blocking buffer (PBS/3% BSA) for 2 hours, the mAb EmG3IgG1 was applied at a dilution of 1:10 in blocking buffer for 1 h, followed by a secondary goat-anti-rabbit 10 nm gold conjugate (Aurion, Wageningen, Holland) diluted 1:5 in blocking buffer for 1 h. After several washes in PBS, specimens were fixed in 2% glutaraldehyde phosphate buffer for 10 min, rinsed in water and air-dried. Contrasting by using lead citrate and uranyle acetate was done as previously described (Winzer et al., 2020 (link)). Specimens were viewed on a Morgagni Transmission Electron Microscope operating at 80 kV.
Lr white acrylic resin
Manufactured by Merck Group
Sourced in Sao Tome and Principe, United States
LR White acrylic resin is a laboratory-grade embedding medium for use in electron microscopy. It is a methacrylate-based resin that is widely used for the preparation of ultrathin sections for transmission electron microscopy (TEM) and scanning electron microscopy (SEM) analysis. The resin provides a stable, uniform matrix for embedding and supporting biological samples during sectioning and imaging.
Automatically generated - may contain errors
Lab products found in correlation
Glutaraldehyde, by Merck Group (2 mentions)
Em uc7 ultramicrotome, by Leica (1 mentions)
Em uc7 microtome, by Leica (1 mentions)
Eclipse ni, by Nikon (1 mentions)
Eclipse ni u, by Nikon (1 mentions)
Sodium nitrite, by Merck Group (1 mentions)
Calcein, by Merck Group (1 mentions)
Vs120 whole slide imager, by Olympus (1 mentions)
Naphthol as bi phosphate substrate, by Merck Group (1 mentions)
Pararosaniline dye, by Merck Group (1 mentions)
Tecnai g2, by Thermo Fisher Scientific (1 mentions)
Diamond knife, by Leica (1 mentions)
Field emission transmission electron microscope, by Thermo Fisher Scientific (1 mentions)
Lead citrate, by Merck Group (1 mentions)
Uranyl acetate, by Merck Group (1 mentions)
Digital micrograph software, by Ametek (1 mentions)
Osmium tetroxide, by Merck Group (1 mentions)
Jem 1400, by JEOL (1 mentions)
Aqueous glutaraldehyde, by Merck Group (1 mentions)
Fuming hydrochloric acid, by Merck Group (1 mentions)
12 protocols using lr white acrylic resin
1
Ultrastructural Localization of EmG3 in E. multilocularis
Check if the same lab product or an alternative is used in the 5 most similar protocols
In vitro cultured E. multilocularis metacestodes were harvested after 6 months, washed in 100 mM sodium phosphate buffer (pH 7.2) and fixed in phosphate buffer containing 3% paraformaldehyde and 0.05% glutaraldehyde for 2 hours at room temperature. They were then washed in PBS, and sequentially dehydrated in ethanol (30, 50, 70, 90 and 100%) and embedded in LR White acrylic resin (Sigma Aldrich) at -20°C, with three changes of resin during 3 days. Polymerization was done at 60°C overnight. Ultrathin sections were cut by using an ultramicrotome (Reichert and Jung, Vienna, Austria) and were placed on 300 mesh formvar-carbon-coated nickel grids (Plano GmbH, Marburg, Germany). After incubating sections on a drop of blocking buffer (PBS/3% BSA) for 2 hours, the mAb EmG3IgG1 was applied at a dilution of 1:10 in blocking buffer for 1 h, followed by a secondary goat-anti-rabbit 10 nm gold conjugate (Aurion, Wageningen, Holland) diluted 1:5 in blocking buffer for 1 h. After several washes in PBS, specimens were fixed in 2% glutaraldehyde phosphate buffer for 10 min, rinsed in water and air-dried. Contrasting by using lead citrate and uranyle acetate was done as previously described (Winzer et al., 2020 (link)). Specimens were viewed on a Morgagni Transmission Electron Microscope operating at 80 kV.
+ Expand
In vitro cultured E. multilocularis metacestodes were harvested after 6 months, washed in 100 mM sodium phosphate buffer (pH 7.2) and fixed in phosphate buffer containing 3% paraformaldehyde and 0.05% glutaraldehyde for 2 hours at room temperature. They were then washed in PBS, and sequentially dehydrated in ethanol (30, 50, 70, 90 and 100%) and embedded in LR White acrylic resin (Sigma Aldrich) at -20°C, with three changes of resin during 3 days. Polymerization was done at 60°C overnight. Ultrathin sections were cut by using an ultramicrotome (Reichert and Jung, Vienna, Austria) and were placed on 300 mesh formvar-carbon-coated nickel grids (Plano GmbH, Marburg, Germany). After incubating sections on a drop of blocking buffer (PBS/3% BSA) for 2 hours, the mAb EmG3IgG1 was applied at a dilution of 1:10 in blocking buffer for 1 h, followed by a secondary goat-anti-rabbit 10 nm gold conjugate (Aurion, Wageningen, Holland) diluted 1:5 in blocking buffer for 1 h. After several washes in PBS, specimens were fixed in 2% glutaraldehyde phosphate buffer for 10 min, rinsed in water and air-dried. Contrasting by using lead citrate and uranyle acetate was done as previously described (Winzer et al., 2020 (link)). Specimens were viewed on a Morgagni Transmission Electron Microscope operating at 80 kV.
2
Immunogold Labeling of Mycobacterium Tuberculosis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For sample preparation, standard protocols were followed with minor modifications [19 (link)–22 (link)]. Briefly, log phase cells of Mtb H37Rv were fixed overnight with in PBS containing 4% paraformaldehyde and 0.2% glutaraldehyde at 4°C. It was then washed twice with 0.1M cacodylate buffer and subjected to dehydration with an ethanol gradient (30%, 50%, 70%, 80% and 90%) for 30 min and absolute ethanol for 1h at 4°C. Embedding was done in LR-white acrylic resin (Sigma-aldrich) for 36h at 55°C. Ultrathin sections (80nm thick) were cut with a Leica EM UC7 ultramicrotome and collected on nickel grids and processed for immunogold labeling. Grids were blocked with PBS containing 0.01% Tween-20 and 2% skimmed milk for 30 min at room temperature followed by three washes in PBST. Primary antibody incubation was done with 1:100 dilution of anti-Cut5b antiserum or pre-immune serum for 16–18 h at 4°C. Grids were washed and incubated with a 1:5 dilution of 10nm gold nanoparticles conjugated goat- anti- mouse secondary antibody (Sigma) for 1h at room temperature. Three washes were performed with PBST and one with water before staining the grids with 2% aqueous uranyl acetate. Samples were then observed under JEOL 2100 TEM.
3
Fixation and Resin Embedding of Plant Seeds
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mature dry seeds of Col-0, abi3-6, gs1 and transgenic plants were fixed in PBS buffer containing 10% formaldehyde, 5% acetic acid, and 50% ethanol and subsequently dehydrated in a (10, 30, 50, 70, 80, 90,100% 1 h each step) graded ethanol series overnight. Seeds were then infiltrated with graded concentrations of LR White acrylic resin (Sigma, Co., St. Louis, MO, United States) at 25, 50, and 75%, each for a minimum of 3 h followed by two changes of 16 h 100% resin. Seeds were placed in fresh 100% LR White acrylic resin in an embedding capsule and polymerized at 60°C for 24 h, then observed under a light microscope (Leica, Germany).
4
Histological and Cytological Observations of Tagetes erecta Anthers
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For histological observations, isolated T. erecta anthers were fixed in a mixture of 2.5% paraformaldehyde and 2.5% glutaraldehyde in cacodylate buffer (pH 7.2) for 24 h at room temperature. The material was then rinsed in cacodylate buffer and placed in a 2% solution of osmium tetraoxide in deionised water. Next, it was dehydrated in a series of alcohol, placed in 100% ethanol, and embedded in LR White acrylic resin (Sigma). The material was cut into semi-thin sections (1 μm) using a Leica EM UC7 microtome (Wetzlar, Germany) and stained with a 1% toluidine blue solution. The sections were prepared under a light microscope Nikon Eclipse Ni-U with a digital camera and NIS-Elements BP software.
Crushed preparations were made for cytological observations. Differently sized anthers of T. erecta were crushed and stained with acetocarmine (Gerlach 1977 ). The observations were carried out under a light microscope Nikon Eclipse Ni with Nomarski contrast. Photographic documentation was made with a digital camera and NIS-Elements BP software.
Crushed preparations were made for cytological observations. Differently sized anthers of T. erecta were crushed and stained with acetocarmine (Gerlach 1977 ). The observations were carried out under a light microscope Nikon Eclipse Ni with Nomarski contrast. Photographic documentation was made with a digital camera and NIS-Elements BP software.
5
Histopathological Analysis of Nematode Parasitism
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Histopathological analysis was set up to evaluate the effect of P. chlamydosporia on nematode parasitism, in particular the effect on the nematode feeding site in the presence and absence of the fungus. Galls and healthy portions of roots of the FN- and N-treated plants were hand-dissected and fixed in a mix of 1.5% glutaraldehyde and 3% paraformaldehyde in 10 mM phosphate-buffered saline (PBS) containing 150 mM NaCl (pH 7.2) for 3 h at room temperature, dehydrated in a graded ethanol series to absolute ethanol and then embedded in LR White acrylic resin (Sigma, St. Louis, MO, USA), left to polymerize overnight at 60 °C. Around 15–20 root samples were taken for each treatment, which were cut by an ultramicrotome to obtain 2.5 μm thick sections. After cutting, the sections were collected on a slide, stained with 1% Toluidine blue solution (1 g Na tetrahydroborate, 1 g of Toluidine blue, 100 mL H2O) and observed under a light microscope.
6
Histology of Plant Nectaries and Trichomes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To obtain semi-thin sections, fragments of ovaries with nectaries and sepals with trichomes (4 × 4 mm, n = 5) were fixed in 2.5% glutaraldehyde in 0.1 M phosphate buffer at pH 7.2 for 12 h at a temperature of 4 °C. Next, they were carefully washed three times in phosphate buffer, dehydrated in an ethanol series, and embedded in LR white resin (LR white acrylic resin, medium grade, Sigma-Aldrich). Semi-thin sections, with a thickness in the range of 70–80 μm, were cut with glass knives using a Reichert Ultracut S ultramicrotome. For general histology, the semi-thin sections were stained with a 1% aqueous methylene blue-azure II solution (O’Brien and McCully 1981 ). The presence of water-insoluble polysaccharides was detected using Periodic acid-Schiff’s (PAS) reagent (O’Brien and McCully 1981 ) after blocking of free aldehyde groups.
7
Histological Analysis of Osteoclasts and Bone Formation
8
Immunogold Localization of Rhodopsin and ETC I in Bovine Retina
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Front halves of bovine eyes were excised and the vitreous humor and lens removed. Eye cups were then filled with fixative consisting of 4% paraformaldehyde and 0.1% glutaraldehyde in PBS buffer solution. After fixation (ON at 4°C), retinas were removed from the eye capsule, cut into small pieces, washed overnight with 50 mmol/L NH4Cl, dehydrated and embedded in LR White Acrylic Resin (Sigma ‐Aldrich, St.louis, MO), and polymerized at 58°C. Ultrathin sections were placed on nickel grids and used the next day for postembedding immunogold experiments. Sections were treated with blocking solution (1% BSA, 0.1% Tween 20, PBS 1×), then incubated with mouse monoclonal anti‐RHO (1:100) (Sigma Aldrich) and rabbit polyclonal anti‐ND1 subunit of ETC I Antibody (Ab) (diluted 1:50) (Abcam) overnight at 4°C. Ab binding was detected using secondary goat anti‐mouse IgG (British BioCell International) (diluted 1:100) coupled to gold particles (5 nm) and anti‐rabbit IgG (British BioCell International) (diluted 1:100) coupled to gold particles (25) nm. Sections were analyzed at a FEI Tecnai G2 transmission electron microscope operating at 100 kV. In negative controls, the preimmune serum was applied to the sections instead of the specific primary Ab. Images were acquired with OSIS Veleta cameras, collected and typeset in Corel Draw X6.
9
Cell-cell Junction Dynamics in Cryptosporidium Infection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To observe cell-cell junctions after C. parvum infection, the ileum samples were selected from a C. parvum IId infected group and control group at 3 dpi, 11 dpi, and 30 dpi. The ileum samples were fixed overnight at 4˚C in 4% paraformaldehyde and 0.1% glutaraldehyde in PBS, washed with PBS, embedded in low-melting agarose and dehydrated in a gradient of ethanol at -20˚C. The samples were then infiltrated with LR White acrylic resin (Sigma, St. Louis, MO, USA) for overnight at -20˚C and polymerized with fresh LR White in gelatin capsules at -25˚C for three days. Thin sections measuring 50 nm were obtained using a diamond knife from Leica (Wetzlar, Germany) and fixed onto nickel grids. The slices were viewed using a field emission transmission electron microscope (FEI Company, Hillsboro, OR, USA).
10
Morphological Analysis of Cold-Adapted Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The morphological changes of cold-adapted cells were analyzed using a transmission electron microscope (JEM-1400; Jeol, Japan). Prior to observation, each sample fixed in 2% glutaraldehyde (Merck, Germany) was post-fixed overnight in 1% osmium tetroxide (Sigma-Aldrich), dehydrated with a graded ethanol series of 30%, 50%, and 70%, embedded in LR white acrylic resin (Sigma-Aldrich) at 50°C for 24 h, and then sectioned using an ultra-microtome with a diamond knife. The sections were placed on the center of a TEM grid and stained with uranyl acetate (Sigma-Aldrich) and lead citrate (Sigma-Aldrich). EPS thickness was examined using DigitalMicrograph software (Gatan Inc., USA). The average diameter was calculated from the values for 16 bacteria taken at random locations of each cell.
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!