Formvar coated nickel grid

Manufactured by Electron Microscopy Sciences

Sourced in United States

Formvar-coated nickel grids are a type of lab equipment used in electron microscopy. They provide a thin, transparent film that supports and stabilizes specimens during imaging. The nickel material offers durability and conductivity for electron beam applications.

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Lab products found in correlation

H7650 tem microscope, by Hitachi (2 mentions) Vivaspin, by Sartorius (2 mentions) Uranyl acetate and lead citrate, by Electron Microscopy Sciences (1 mentions) Epoxy resin, by Electron Microscopy Sciences (1 mentions) Glutaraldehyde, by Electron Microscopy Sciences (1 mentions) Lr white acrylic resin, by Electron Microscopy Sciences (1 mentions) Em uc6 ultramicrotome, by Leica (1 mentions) Talos l120c, by Thermo Fisher Scientific (1 mentions) Jem 1010 transmission electron microscope, by JEOL (1 mentions) Ht7700 tem, by Hitachi (1 mentions) Jem 1011 transmission electron microscope, by JEOL (1 mentions)

12 protocols using formvar coated nickel grid

Transmission and Scanning Electron Microscopy of Extracellular Vesicles

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For TEM analysis, several drops of EV suspension (20 μl/drop) were placed on Parafilm. Formvar-coated nickel grids (Electron Microscopy Sciences, Hatfield, Pa, USA) were gently placed on the top of each drop for 60 minutes at room temperature, the coated side of the grid facing the suspension. After washing on phosphate buffer (PB) 0.1 M pH 7.3, grids were fixed with 2.5% glutaraldehyde (Fluka, St. Louis, MO, USA) for 5 minutes, washed in distilled water, and contrasted with 2% aqueous uranyl acetate. Finally, the grids were washed in distilled water, air dried, and observed under a Philips EM 208 transmission electron microscope equipped with a digital camera (University Centre for Electron Microscopy (CUME) Perugia).
For SEM analysis, EVs were allowed to adhere to Formvar-coated nickel grids and fixed with 2.5% glutaraldehyde as described for TEM. The grids were attached on metal stubs, coated with chrome to a thickness of 10 nm, and examined with a ZEISS LEO 1525 (Nanomaterials Laboratory, University of Perugia).

Capomaccio S., Cappelli K., Bazzucchi C., Coletti M., Gialletti R., Moriconi F., Passamonti F., Pepe M., Petrini S., Mecocci S., Silvestrelli M, & Pascucci L. (2019). Equine Adipose-Derived Mesenchymal Stromal Cells Release Extracellular Vesicles Enclosing Different Subsets of Small RNAs. Stem Cells International, 2019, 4957806.

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Ultrastructural Characterization of Brown Adipose Tissue

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For ultrastructural characterization by transmission electron microscopy (TEM), interscapular brown adipose tissue sections were fixed with 2% glutaraldehyde (Electron Microscopy Sciences, Hatfield, PA, USA) and immediately dehydrated through a series of graded ethanol dilutions. Samples were embedded in LR White acrylic resin or epoxy resin (Electron Microscopy Sciences). Ultrathin sections (80-nm-thick) cut with a Leica EM UC6 ultramicrotome (Leica Microsystems Canada, Ltd, Richmond Hill, ON) were placed on formvar-coated nickel grids (Electron Microscopy Sciences) and stained (or left unstained) with uranyl acetate and lead citrate (Electron Microscopy Sciences) for viewing by TEM. A field-emission FEI Tecnai 12 BioTwin TEM (FEI, Hillsboro, OR, USA) was used to image the stained sections at 120 kV.
Electron diffraction in the selected-area configuration (SAED) mode, and energy-dispersive X-ray spectroscopy (EDS), were performed on unstained sections at 200 kV using a Philips CM200 TEM equipped with a Gatan Ultrascan 1000 2k X 2 k CCD camera system model 895 and an EDAX Genesis EDS analysis system (FEI, Hillsboro, OR, USA).

Herrmann M., Babler A., Moshkova I., Gremse F., Kiessling F., Kusebauch U., Nelea V., Kramann R., Moritz R.L., McKee M.D, & Jahnen-Dechent W. (2020). Lumenal calcification and microvasculopathy in fetuin-A-deficient mice lead to multiple organ morbidity. PLoS ONE, 15(2), e0228503.

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Nanoparticle Negative Staining Protocol

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Nanoparticle sample solutions were transferred to an ammonium acetate buffer solution and then negatively stained using an ammonium acetate buffer containing 2% sodium phosphotungstate. Drops of samples were then placed onto 100 mesh Formvar-coated nickel grids (Electron Microscopy Sciences) and allowed to air-dry. Grids were imaged using a Hitachi 7650 microscope operated at 80 kV and connected to a digital camera (Scientific Instruments and Applications) controlled by Maxim CCD software.

Fay F., Hansen L., Hectors S.J., Sanchez-Gaytan B.L., Zhao Y., Tang J., Munitz J., Alaarg A., Braza M.S., Gianella A., Aaronson S.A., Reiner T., Kjems J., Langer R., Hoeben F.J., Janssen H.M., Calcagno C., Strijkers G.J., Fayad Z.A., Pérez-Medina C, & Mulder W.J. (2017). Investigating the Cellular Specificity in Tumors of a Surface-Converting Nanoparticle by Multimodal Imaging. Bioconjugate chemistry, 28(5), 1413-1421.

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Transmission Electron Microscopy of Extracellular Vesicles

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The morphology of EVs from UFs was determined using transmission electron microscopy (TEM; Talos L120C, Thermo Fisher) based on a previously described method [34 (link),35 (link)]. In brief, the re-suspended EVs were loaded onto Cu grids and incubated at room temperature for 10 min. Then, the grids were stained with 2% uranyl acetate for 2 min and dried overnight at room temperature.
TEM analysis was also used to assess the presence of EVs in the endometrium by modification of a previously described method [36 ]. Briefly, tissues were dehydrated in alcohol, embedded in epoxy resin, ultra-sectioned (less than 1 mm³), and transferred to 300-mesh Formvar coated nickel grids (Electron Microscopy Sciences). A mixture of 4% uranyl acetate and lead citrate was then added to the sections for 15 min, and the grids were dried overnight at room temperature. All 6 tissue samples (C16, n = 3; C16, n = 3) were analyzed for TEM. The grids were then observed under a transmission electron microscope and the images were analyzed.

Xie Y., Liu G., Zang X., Hu Q., Zhou C., Li Y., Liu D, & Hong L. (2021). Differential Expression Pattern of Goat Uterine Fluids Extracellular Vesicles miRNAs during Peri-Implantation. Cells, 10(9), 2308.

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Transmission Electron Microscopy Analysis of EVs

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To assess the presence of EVs in the endometrium and CAM, standard TEM procedure was used. Briefly, tissues were dehydrated in alcohol, embedded in epoxy resin, ultrasectioned, transferred to 300-mesh Formvar coated nickel grids (Electron Microscopy Sciences) and stained using 4% uranyl acetate and lead citrate according to previously published methods45 (link). The grids were subsequently observed using a JEM-1010 transmission electron microscope (Jeol, Korea) at 100 kV. Images were obtained and processed using Gatan Microscopy Suite (GMS) 3 Software (Gatan, Inc. Pleasanton, CA, USA).

Bidarimath M., Khalaj K., Kridli R.T., Kan F.W., Koti M, & Tayade C. (2017). Extracellular vesicle mediated intercellular communication at the porcine maternal-fetal interface: A new paradigm for conceptus-endometrial cross-talk. Scientific Reports, 7, 40476.

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Nanoparticle Morphology Imaging by TEM

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The nanoparticles were loaded onto formvar-coated nickel grids (Electron Microscopy Sciences), followed by negative staining using a 2% phosphotungstic acid solution (Sigma). The morphology of the nanoparticles was imaged using HT7700 TEM (Hitachi) at 120 kV, coupled with a digital micrograph camera and software suite (Gatan).

Wolski V.M., Soltyk A.M, & Brunton J.L. (2001). Mouse Toxicity and Cytokine Release by Verotoxin 1 B Subunit Mutants. Infection and Immunity, 69(1), 579-583.

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Ultrastructural Analysis of Mouse Sperm

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Mouse sperm bound to beads were fixed in 2% glutaraldehyde in 0.1 M cacodylate buffer (pH 7.4) and incubated at 4°C for 2 hours. After extensive washing in the cacodylate buffer, beads were embedded in 2% agarose. The samples were then dehydrated through a graded series of ethanol and processed for embedding in LR White resin. Ultrathin sections were obtained with an ultramicrotome (Microm International GmbH) and mounted on formvar-coated nickel grids (Electron Microscopy Sciences). Ultrathin sections were counterstained with uranyl acetate followed by lead citrate and imaged in a Jeol JEM-1011 transmission electron microscope (Jeol).

Avella M.A., Baibakov B.A., Jimenez-Movilla M., Sadusky A.B, & Dean J. (2016). ZP2 peptide beads select human sperm in vitro, decoy mouse sperm in vivo, and provide reversible contraception. Science translational medicine, 8(336), 336ra60.

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Visualizing Aβ42 Aggregation Modulation

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We observed the effect of α-TOC, α-T3, and γ-T3 on Aβ42 aggregation and disaggregation with SEM imaging. We carried out sample preparation according to a previous study with modification [10 (link)]. Briefly, we spotted the reaction mixture onto a 150 mesh Formvar-coated nickel grid (Electron Microscopy Sciences, Hatfield, PA, USA) and dried for 20 min at room temperature. Then, we displaced the sample with 2.5% v/v glutaraldehyde in water for 5 min. We then washed the grid by dipping it in water 5 times, repeating the washing step 3 times. Next, we stained the grid with 2% filtered uranyl acetate in water for 30 s, further washed it 3 times in water, and allowed it to dry at room temperature. We observed samples using a field emission SEM (JEOL JSM-7505FA; JEOL, Peabody, MA, USA) at 25 kV. We selected the images of SEM randomly from the 150 mesh Formvar-coated nickel grid that was used to place the reaction mixtures, at a magnification of 25000X.

Ibrahim N.F., Hamezah H.S., Yanagisawa D., Tsuji M., Kiuchi Y., Ono K, & Tooyama I. (2021). The effect of α-tocopherol, α- and γ-tocotrienols on amyloid-β aggregation and disaggregation in vitro. Biochemistry and Biophysics Reports, 28, 101131.

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TEM Imaging of Nanoparticles in Acetate Buffer

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The nanoparticle’s original buffer solution was exchanged with acetate buffer (0.125 M ammonium acetate, 2.6 mM ammonium carbonate, and 0.26 mM tetrasodium ethylenediaminetetraacetate (EDTA) at pH 7.4) using centrifugal concentration devices (MWCO 100 000). A 10 μL solution of nanoparticles in acetate buffer was mixed with 10 μL of a 2% phosphotungstic acid solution to be negatively stained and then cast on a 100 mesh Formvar coated nickel grid (Electron Microscopy Sciences). The TEM images were acquired using a Hitachi 7650 TEM microscope operated at 80 kV coupled to a Scientific Instruments and Applications (SIA) digital camera controlled by Maxim CCD software.

Sanchez-Gaytan B.L., Fay F., Lobatto M.E., Tang J., Ouimet M., Kim Y., van der Staay S.E., van Rijs S.M., Priem B., Zhang L., Fisher E.A., Moore K.J., Langer R., Fayad Z.A, & Mulder W.J. (2015). HDL-Mimetic PLGA Nanoparticle To Target Atherosclerosis Plaque Macrophages. Bioconjugate chemistry, 26(3), 443-451.

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Negative Staining of HDL Particles

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TEM microscopy images of selected samples were acquired using a Hitachi H-7650 TEM microscope (Hitachi High Technologies, Pleasanton, CA) operating at 80 kV, coupled to a Scientific Instruments and Applications digital camera, controlled by the Maxim charge-coupled device software.
HDL samples were centrifuged in (MWCO 100 000) centrifugal concentration tubes (Vivaspin; Sartorius Corporation, Edgewood, NY) to replace the original buffer solution by acetate buffer (0.125 M ammonium acetate, 2.6 mM ammonium carbonate, and 0.26 mM tetrasodium ethylenediaminetetraacetate at pH 7.4). Afterward, 10 μL of HDL solution was mixed with 10 μL of 2% phosphotungstic acid for negative staining and casted on a 100-mesh Formvar-coated nickel grid (Electron Microscopy Sciences, Hatfield, PA).

Ramos-Cabrer P., Fay F., Sanchez-Gaytan B.L., Tang J., Castillo J., Fayad Z.A, & Mulder W.J. (2016). Conformational Changes in High-Density Lipoprotein Nanoparticles Induced by High Payloads of Paramagnetic Lipids. ACS Omega, 1(3), 470-475.

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