Formvar and carbon coated copper grid

Manufactured by Ted Pella

Sourced in United States

Formvar and carbon-coated copper grid is a type of laboratory equipment used for sample preparation in electron microscopy. It consists of a copper grid coated with a thin layer of Formvar (a polyvinyl formal resin) and carbon, which provides a stable and conductive surface for mounting and analyzing samples.

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Lab products found in correlation

Tecnai g2 spirit biotwin, by Thermo Fisher Scientific (1 mentions) 1 100 transmission electron microscope tem, by JEOL (1 mentions) Cm12 electron microscope, by TVIPS (1 mentions) 1kx1k ccd camera, by TVIPS (1 mentions) Vitrobot mk 4, by Thermo Fisher Scientific (1 mentions) Fe tem, by JEOL (1 mentions) H 7650 transmission electron microscope, by Hitachi (1 mentions) Uranyl acetate, by Polysciences (1 mentions) Em 400 transmission electron microscope, by Philips (1 mentions) Smartlab x ray diffractometer, by Rigaku (1 mentions)

Close spelling variants of this product

Formvar and carbon coated 200-mesh copper grid Formvar/carbon-coated copper grids Carbon-Formvar-coated copper grids Formvar-coated copper grid Formvar carbon-coated grid Carbon-coated copper grids Carbon/formvar grids Formvar/carbon grid Carbon-coated grids Formvar and carbon

8 protocols using formvar and carbon coated copper grid

Exosome Visualization by Transmission Electron Microscopy

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The cells to be observed were fixed with 2.5% normal glutaraldehyde in PBS, aspirated into the centrifuge tube, and centrifugated at 2000 rpm for 2 min. The fixative was discarded followed by a new fixative for electron microscopy, and the cells were observed under TEM (Hitachi).
Exosome samples were transferred into 200-mesh Formvar and carbon-coated copper grid (Ted Pella) and incubated for 1 min. After adsorption, the grid was rinsed with water, and the excess solution was removed. Grids were then stained with 1% phosphotungstic acid for 1 min, and the excess solution was cleared. Exosomes were observed by TEM (FEI Tecnai G2 Spirit BioTWIN) after drying at room temperature.

Ni Z., Li Y., Song D., Ding J., Mei S., Sun S., Cheng W., Yu J., Zhou L., Kuang Y., Li M., Cai Z, & Yu C. (2022). Iron-overloaded follicular fluid increases the risk of endometriosis-related infertility by triggering granulosa cell ferroptosis and oocyte dysmaturity. Cell Death & Disease, 13(7), 579.

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Extracellular Vesicle Isolation from UC-MSC

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After harvesting UC-MSCs via either the manual manufacturing process or the automated culture platform, the cell culture supernatants were collected for extracellular vesicle (EV) collection using the ultracentrifugation method as previously described [53 ]. Briefly, the supernatants were centrifuged at 300×g for 10 min at 4 °C to remove the cell pellet. The collected supernatants were then centrifuged at 2000×g for 20 min at 4 °C to remove dead cells and large EVs and then at 10,000×g for 30 min at 4 °C to remove cell debris. The supernatants were then centrifuged at 100,000×g for 70 min at 4 °C to collect pellets containing EVs and contaminating proteins. The pellets were washed with 10 mL PBS and subjected to an additional centrifugation at 100,000×g for 70 min at 4 °C to remove contaminating protein. The remaining EVs were resuspended in 100 μl PBS and stored at −80 °C for further use. To visualize and measure the EV size, TEM images of EV samples were obtained using a JEOL 1100 Transmission Electron Microscope (TEM, JEOL Ltd., Tokyo, Japan) at 80 kV. For visualization, EV samples were dropped and dried on a formvar- and carbon-coated copper grid (Ted Pella, Inc.).

Hoang D.M., Nguyen Q.T., Phan T.T., Ngo A.T., Pham P.T., Bach T.Q., Le P.T., Bui H.T, & Thanh L.N. (2023). Advanced cell-based products generated via automated and manual manufacturing platforms under the quality by design principle: Are they equivalent or different?. Heliyon, 9(5), e15946.

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Transmission Electron Microscopy of Biological Samples

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All electron microscopy samples were fixed in 2% (vol/vol) glutaraldehyde (EM grade, purchased from EMS, Hatfield, PA, USA) directly in the growth medium for several hours and then washed in phosphate buffered saline (PBS). For Transmission Electron Microscopy (TEM), a 5 μl sample was put onto a formvar and carbon coated copper grid (200 mesh, Ted Pella, Redding, CA, USA), which was freshly glow-discharged in order to make the carbon film hydrophilic. The sample was allowed to settle for 5 min and the liquid removed with filter paper. Immediately 5 μ l of a 2% (wt/vol) aqueous solution of uranyl acetate was put onto the grid and left for 1 min before also being dried with filter paper. Two quick washes (10 μl each) with distilled water followed. After drying, the grids were investigated with a Phillips Tecnai 12 electron microscope (FEI Company, Hillsboro, OR, USA) with a 120 kV accelerating voltage and magnifications typically between 2900 × and 9300 ×. A Gatan camera (Gatan, Pleasanton, CA, USA) was used for image acquisition.

Ray J., Keller K.L., Catena M., Juba T.R., Zemla M., Rajeev L., Knierim B., Zane G.M., Robertson J.J., Auer M., Wall J.D, & Mukhopadhyay A. (2014). Exploring the role of CheA3 in Desulfovibrio vulgaris Hildenborough motility. Frontiers in Microbiology, 5, 77.

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Transmission Electron Microscopy of Extracellular Vesicles

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For TEM, EV samples at ∼2.0E+11 p/mL were transferred to a 200-mesh Formvar and carbon-coated copper grid (Ted Pella) and incubated for 1 min. After adsorption, the grids were rinsed with water and the excess solution was wicked away. The grids were then stained with a 1% solution of uranyl acetate for ∼30 s. After staining, the excess solution was wicked away and the grids were allowed to dry. The negatively stained samples were analyzed with a FEI Morgagni (Hillsboro) electron microscope operating at 80 kV.
For cryo-TEM, EV samples at 2.0E+12 p/mL were transferred to a glow-discharged R2/2 Quantifoil Holey carbon film grid and incubated for 1 min. Grids were blotted and immediately plunge frozen into nitrogen-cooled liquid ethane using a Vitrobot Mk IV (ThermoFisher). The samples were imaged at 13,000× magnification using a Phillips CM12 electron microscope operated at 100 keV and captured with a TVIPS 1kx1k CCD camera.

Dooley K., McConnell R.E., Xu K., Lewis N.D., Haupt S., Youniss M.R., Martin S., Sia C.L., McCoy C., Moniz R.J., Burenkova O., Sanchez-Salazar J., Jang S.C., Choi B., Harrison R.A., Houde D., Burzyn D., Leng C., Kirwin K., Ross N.L., Finn J.D., Gaidukov L., Economides K.D., Estes S., Thornton J.E., Kulman J.D., Sathyanarayanan S, & Williams D.E. (2021). A versatile platform for generating engineered extracellular vesicles with defined therapeutic properties. Molecular Therapy, 29(5), 1729-1743.

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TEM Imaging of Alum and PGA/Alum

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TEM images of alum and PGA/Alum were obtained using a field-emission transmission electron microscope (FE-TEM; JEOL Ltd.). For visualization, alum and PGA/Alum (100 μg/ml) solutions were dropped and dried on a formvar- and carbon-coated copper grid (Ted Pella, Inc).

Nguyen Q.T., Kwak C., Lee W.S., Kim J., Jeong J., Sung M.H., Yang J, & Poo H. (2019). Poly-γ-Glutamic Acid Complexed With Alum Induces Cross-Protective Immunity of Pandemic H1N1 Vaccine. Frontiers in Immunology, 10, 1604.

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Visualizing Microparticles with TEM

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To visualise microparticles using transmission electron microscopy (TEM), 5 µL from each sample fraction was placed on a 200-mesh formvar and carbon coated copper grid (Ted Pella, Redding, CA, USA) for 10 min. Excess samples were wicked away before the grid was floated on a 20 μL drop of 25% Uranyl Acetate Alternative (Ted Pella) diluted in distilled water for 1 min. Excess stain was wicked away to yield a dry grid prior to image acquisition using a Hitachi H7650 transmission electron microscope (Maidenhead, UK) operating at 100 kV and side mounted camera.

Hu S., Cavanagh B.L., Harrington R., Ahmad M., Kearns G., Meaney S, & Wynne C. (2020). A Novel Pool of Microparticle Cholesterol Is Elevated in Rheumatoid Arthritis but Not in Systemic Lupus Erythematosus Patients. International Journal of Molecular Sciences, 21(23), 9228.

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Phage Purification and Characterization Protocol

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A total of 1 mL of phages (titer 2 × 10⁹ PFU/mL) was centrifuged at 24,500× g for 3 h at 4 °C. Subsequently, the obtained pellet was washed twice with a 5% ammonium molybdate solution (Sigma-Aldrich, St. Louis, MO, USA), pH 6.0, and was suspended in 5% ammonium molybdate to the titer of 10¹¹ PFU/mL. Next, a drop of the suspension was placed onto a formvar and carbon-coated copper grid (TED Pella, Inc., Redding, CA, USA) and was allowed to absorb the sample on the filter paper. The phages were stained with 2% uranyl acetate (Polysciences, Inc., Warrington, PA, USA) for 1 min and were observed under a transmission electron microscope (TEM), JEOL 1010 TEM, at 80 kV under the magnification of 60,000–100,000×. Phages were measured (15 virions of phage 2e and 18 virions of phage 3e) using the NIS-Elements D 5.11.01 software. The identification and classification of phages were conducted according to the guidelines of the International Committee on the Taxonomy of Viruses (ICTVs). Phages were classified using Ackermann’s viral specifications [31 ].

Moryl M., Palatyńska-Ulatowska A., Maszewska A., Grzejdziak I., Dias de Oliveira S., Pradebon M.C., Steier L., Różalski A, & Poli de Figueiredo J.A. (2022). Benefits and Challenges of the Use of Two Novel vB_Efa29212_2e and vB_Efa29212_3e Bacteriophages in Biocontrol of the Root Canal Enterococcus faecalis Infections. Journal of Clinical Medicine, 11(21), 6494.

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Characterization of Nanoparticle Samples

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Transmission electron microscope (TEM) images were collected using a Phillips EM-400 transmission electron microscope operating at an accelerating voltage of 120 kV. Samples were prepared by casting one drop of dilute dispersed sample in toluene onto a 300-mesh Formvar and carbon-coated copper grid (Ted Pella, Inc.). Particle counting analysis used a minimum of 200 individual particles and size was determined using the ImageJ program (http://rsbweb.nih.gov/ij/). Powder X-Ray diffraction (XRD) patterns were collected with a Rigaku SmartLab X-ray Diffractometer in the Bragg-Brentano geometry using Cu Kα radiation. Samples were dropcast in toluene onto either a zero background plate or a fused quartz substrate. Simulated powder XRD patterns were made using the CrystalMaker software suite. Scanning electron microscope (SEM) images of dropcast films were obtained using a Zeiss NVision 40 field emission SEM operating at 3 kV. X-Ray photoelectron spectroscopy (XPS) analyses were performed on a Kratos Axis Ultra X-ray photoelectron spectrometer with a monochromatic Al Kα excitation source operating at 15 kV and 10 mA. Samples were prepared by dropcasting the sample onto a gold-coated Si wafer.

Alberding B.G., Biacchi A.J., Walker A.R, & Heilweil E.J. (2016). Charge Carrier Dynamics and Mobility Determined by Time-Resolved Terahertz Spectroscopy on Films of Nano-to-Micrometer-Sized Colloidal Tin(II) Monosulfide. The journal of physical chemistry. C, Nanomaterials and interfaces, 120(88), 15395-15406.

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