Lacey carbon film

Manufactured by Ted Pella

Sourced in United States

Lacey carbon film is a type of thin, perforated carbon film used as a support medium for transmission electron microscopy (TEM) specimen preparation. The film features randomly distributed holes, providing an open structure that allows the electron beam to pass through the specimen during TEM analysis.

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Lab products found in correlation

Jem 2100f, by JEOL (2 mentions) Jem 1400, by Ametek (1 mentions) Jem 1400 transmission electron microscope, by JEOL (1 mentions) Ultima 4, by Rigaku (1 mentions) Vitrobot mark 4, by Thermo Fisher Scientific (1 mentions) Gatan 626 cryo holder, by Ametek (1 mentions) Pelco easiglow, by Ted Pella (1 mentions) Jem 1400plus, by JEOL (1 mentions)

7 protocols using lacey carbon film

TEM Imaging of Nanomaterials

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TEM images were
acquired with a JEOL JEM2100F (JEOL Ltd.) microscope operating at
200 kV. Each sample was prepared by drop-casting on 400 mesh Cu grids
coated with a lacey carbon film (Ted Pella, Inc.) and dried overnight
under vacuum at room temperature.

Karadaghi L.R., To A.T., Habas S.E., Baddour F.G., Ruddy D.A, & Brutchey R.L. (2022). Activating Molybdenum Carbide Nanoparticle Catalysts under Mild Conditions Using Thermally Labile Ligands. Chemistry of Materials, 34(19), 8849-8857.

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Visualizing Extracellular Membrane Vesicles

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EMVs were visualized by TEM as previously reported (Yokoyama et al., 2017 (link)). Two microliters of the EMV samples were adsorbed onto hydrophilized carbon-coated copper grids and then treated twice with 2% uranyl acetate to negatively stain the samples. The TEM images were obtained with a JEM-1400 transmission electron microscope (JEOL, Ltd., Tokyo, Japan) at an accelerating voltage of 120 kV. Images were acquired using a charge-coupled device (CCD) camera (a built-in camera in the JEM-1400).
The intact structure of EMVs in buffer was visualized by cryo-electron microscopy (cryo-EM) at −175°C with a side-entry type cryo-specimen holder (Model 626.DH holder, Gatan Inc., Pleasanton, CA, United States). The samples were loaded onto microgrids (lacey carbon film, Ted Pella Inc., Redding, CA, United States) and treated using the same procedures as described previously (Yokoyama et al., 2017 (link)).

Chen C., Kawamoto J., Kawai S., Tame A., Kato C., Imai T, & Kurihara T. (2020). Isolation of a Novel Bacterial Strain Capable of Producing Abundant Extracellular Membrane Vesicles Carrying a Single Major Cargo Protein and Analysis of Its Transport Mechanism. Frontiers in Microbiology, 10, 3001.

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Characterization of Platinum Nanoparticles

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Transmission electron microscopy images were obtained using a JEOL JEM2100F (JEOL Ltd.) microscope operating at 200 kV. Samples were prepared on 400 mesh Cu grid coated with a lacey carbon film (Ted Pella, Inc.) by drop-casting a dilute suspension of PtNPs in ethanol. The size distribution of the PtNPs was determined by analysing 500 unique nanoparticles. Powder X-ray diffraction patterns were collected on a Rigaku Ultima IV diffractometer functioning at 40 mA and 40 kV with a Cu Kα X-ray source (λ=1.5406 Å). The step size and collection time were 0.015° and 10 s per step, respectively. ¹H and ¹⁹F NMR spectra were obtained on Varian 600 spectrometer (600 and 564 MHz, respectively) with chemical shifts reported in p.p.m.

Riche C.T., Roberts E.J., Gupta M., Brutchey R.L, & Malmstadt N. (2016). Flow invariant droplet formation for stable parallel microreactors. Nature Communications, 7, 10780.

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Cryo-TEM Imaging of Peptide Assemblies

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Images for conventional
and cryo-TEM were obtained using a Hitachi HT-7700 Biological TEM
(Hitachi High Technologies America, Schaumburg, IL) equipped with
a LaB₆ filament working at an accelerating voltage of 100
kV. For Cryo-TEM, PA samples were plunged frozen using a Vitrobot
Mark IV (FEI, Hillsboro, OR) operating at 25 °C with 100% humidity.
The PA sample (8 μL) was deposited on 300 square mesh copper
grids with a lacey carbon film (Ted Pella, Redding, CA), blotted,
and plunged into a liquid ethane reservoir cooled by liquid nitrogen.
Following vitrification, the sample was transferred to a Gatan 626
cryo-holder (Gatan, Pleasanton, CA) under liquid nitrogen with the
aid of a transfer stage. Images were acquired using an Orius SC 1000A
CCD camera. All PA formulations were imaged at concentrations of 500
μM in 25 mM HEPES at pH 7.4 buffer. For conventional TEM, PA
samples were dried on carbon film 300 square mesh copper grids (Ted
Pella, Redding, CA) and stained with 0.5% uranyl acetate (UA) solution.
Banding pattern was analyzed using ImageJ by measuring the pixel value
of the gray scaled microscopic images, where a value of 0 represents
white, and a value of 200 or above represents black.

Rubert Pérez C.M., Álvarez Z., Chen F., Aytun T, & Stupp S.I. (2017). Mimicking the Bioactivity of Fibroblast Growth Factor-2 Using Supramolecular Nanoribbons. ACS Biomaterials Science & Engineering, 3(9), 2166-2175.

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Negative-staining Electron Microscopy of VLPs

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Negative-staining electron microscopy of VLPs was adapted from Semionatto et al.²⁴ (link) The 4-1BBL + OX40L VLPs were fixed in 2% uranyl acetates and transferred to 400-mesh copper grids coated by an ultra-thin carbon film superimposed on a Lacey carbon film (Ted Pella, Redding, CA, USA). Images were acquired through a transmission electron microscope (PELCO easiGlow, Ted Pella) equipped with tungsten filament and operated at 120 kV and analyzed using Gatan Digital Micrograph (GMS3) software and ImageJ Fiji software (ImageJ, National Institutes of Health, Rockville, MD, USA).

Palameta S., Manrique-Rincón A.J., Toscaro J.M., Semionatto I.F., Fonseca M.C., Rosa R.S., Ruas L.P., Oliveira P.S, & Bajgelman M.C. (2022). Boosting antitumor response with PSMA-targeted immunomodulatory VLPs, harboring costimulatory TNFSF ligands and GM-CSF cytokine. Molecular Therapy Oncolytics, 24, 650-662.

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Cryogenic Preservation of Neuronal Cultures

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Gold EM grids (200 mesh; with ~97 μm grid hole size) with lacey carbon film (Cat. #: 01882G; Ted Pella INC, Redding, CA, USA) were coated in the same way as the coverslips described in the prior section. The size of most of the holes in the lacey carbon film was between 0.25 and 10 μm. Hippocampal or cortical mouse neurons were cultured on these gold-coated EM grids. At 10–12 DIV, neurons growing on the grids were washed with PBS. Extra liquid was carefully blotted away from the sides of the EM grids where neurons were not adhered using a piece of filter paper. The grids were immediately flash frozen using a homemade manual Cryo-plunger. The frozen grids were transferred and stored at the temperature of liquid nitrogen in a storage tank.

Ma D., Deng B., Sun C., McComb D.W, & Gu C. (2022). The Mechanical Microenvironment Regulates Axon Diameters Visualized by Cryo-Electron Tomography. Cells, 11(16), 2533.

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Transmission Electron Microscopy of EVs

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The EVs preparations were fixed in 2% uranyl acetates and arranged in 400 mesh copper grids coated by an ultra-thin carbon film superimposed on a Lacey carbon film (Ted Pella, Inc., USA). The grids were previously submitted to luminescent discharge of 15 mA/25 s.
The negative staining samples were visualized on the transmission electron microscope (JEOL JEM-1400 Plus) equipped with tungsten filament and operated at 120 kV. The images were analyzed in Gatan Digital Micrograph (GMS3) software.

Semionatto I.F., Palameta S., Toscaro J.M., Manrique-Rincón A.J., Ruas L.P., Paes Leme A.F, & Bajgelman M.C. (2020). Extracellular vesicles produced by immunomodulatory cells harboring OX40 ligand and 4-1BB ligand enhance antitumor immunity. Scientific Reports, 10, 15160.

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