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Epon 812

Manufactured by Electron Microscopy Sciences
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Sourced in United States, United Kingdom

Epon 812 is a low viscosity epoxy resin used for embedding samples for electron microscopy. It provides a hard, durable embedding medium for ultra-thin sectioning.

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Lab products found in correlation

Lead citrate, by Electron Microscopy Sciences (4 mentions) Jem 1011, by JEOL (4 mentions) Glutaraldehyde, by Electron Microscopy Sciences (4 mentions) Uranyl acetate, by Electron Microscopy Sciences (3 mentions) Embed 812, by Electron Microscopy Sciences (3 mentions) Axiophot microscope, by Zeiss (3 mentions) Ultramicrotome, by Leica camera (3 mentions) Pelco silicone rubber molds, by Ted Pella (3 mentions) Tecnai 10 electron microscope, by Philips (3 mentions) Digital imaging system, by Ametek (2 mentions) Cacodylate buffer, by AppliChem (2 mentions) Reichert ultramicrotome, by Leica Microsystems (2 mentions) Morgagni 268d transmission electron microscope, by Thermo Fisher Scientific (2 mentions) Em400 electron microscope, by Philips (2 mentions) Leica ultracut r, by Leica camera (2 mentions) Jem 1400, by JEOL (2 mentions) Osmium tetroxide, by Electron Microscopy Sciences (2 mentions) Axioskop 2 plus, by Zeiss (2 mentions) Ht7700 transmission electron microscope, by Hitachi (2 mentions) Jem 1400plus, by JEOL (2 mentions)

76 protocols using epon 812

1

Ultrastructural Observation of Cell Monolayers

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For ultrastructural observation, cell monolayers were fixed in 2.5% glutaraldehyde (Electron Microscopy Science, Pennsylvania, USA) and in 0.1% cacodylate buffer (Electron Microscopy Science), postfixed in 1% osmium tetroxide (Electron Microscopy Science) and then treated with 1% tannic acid (Acros Organics, New Jersey, NJ, USA). Dehydrated in ethanol (Carlo Erba), cells were detached from the plastic dish by a brief treatment in 1% toluene (Carlo Erba). The pelleted monolayer was then incubated in a solution of 50% toluene and 50% EPON 812 (Electron Microscopy Science) and finally processed for conventional EPON 812 embedding. Ultra-thin sections were cut in a Leica Ultracut R ultramicrotome, contrasted for 10 min in 1% acid tannic and successively in 1% lead hydroxide (Società Italiana Chimici, Rome, Italy) and then viewed in a Hitachi 7000 transmission electron microscope.
Marini E.S., Giampietri C., Petrungaro S., Conti S., Filippini A., Scorrano L, & Ziparo E. (2014). The endogenous caspase-8 inhibitor c-FLIPL regulates ER morphology and crosstalk with mitochondria. Cell Death and Differentiation, 22(7), 1131-1143.
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2

Ultrastructural Analysis of HaCaT Cells

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HaCaT pMSG E5 and HaCaT pMSG cells treated with Dex and stimulated with KGF for 24 h as above were washed three times in PBS and fixed with 2% glutaraldehyde in PBS for 2 h at 4 °C. Samples were postfixed with 1% osmium tetroxide in veronal acetate buffer (pH 7.4) for 1 h at 25 °C, stained with uranyl acetate (5 mg/ml) for 1 h at 25 °C, dehydrated in acetone and embedded in Epon 812 (EMbed 812, Electron Microscopy Science, Hatfield, PA, USA). Ultrathin sections were examined unstained or poststained with uranyl acetate and lead hydroxide, under a Morgagni 268D transmission electron microscope (FEI, Hillsboro, OR, USA) equipped with a Mega View II charge-coupled device camera (SIS, Soft Imaging System GmbH, Munster, Germany) and analyzed with AnalySIS software (SIS).
Belleudi F., Nanni M., Raffa S, & Torrisi M.R. (2015). HPV16 E5 deregulates the autophagic process in human keratinocytes. Oncotarget, 6(11), 9370-9386.
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3

Transmission Electron Microscopy of Mitochondria

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Cells were fixed in 1/2 strength Karnovsky's (2% paraformaldehyde/2.5% glutaraldehyde buffered with 0.2 M cacodylate) and postfixed in 2% OsO4 buffered in 0.2 M cacodylate buffer. After quick dehydration, cells were embedded in Epon 812 (Electron Microscopy Sciences), thin sectioned (70 nm), and stained with uranyl acetate for 2 hours and lead citrate for 5 minutes. The samples were imaged using a JOEL 1230 transmission electron microscope set to 80 kV, and the images were captured using a Gatan digital imaging system. Image analysis was performed using NIH Image J, for mitochondrial size and relative abundance. The latter was calculated by fraction of mitochondria area/cell area.
Dai D.F., Danoviz M.E., Wiczer B., Laflamme M.A, & Tian R. (2017). Mitochondrial Maturation in Human Pluripotent Stem Cell Derived Cardiomyocytes. Stem Cells International, 2017, 5153625.
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4

Evaluating Regenerated Nerve Tissue

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At 6, 12, and 18 weeks post operatively, the regenerated nerves were harvested after SD rats were sacrificed. Specimens of nerves were fixed by 4% paraformaldehyde at room temperature for 24 h. They were washed by PBS for three times. Then the nerve segments were fixed by 1% osmium tetroxide, dehydrated, and embedded in Epon812 (Electron Microscopy Sciences, USA) resin. The cross sections were cut at 4 mm thick (Leica EM UC 6 ultramicrotome) and mounted on gelatin pre-coated slides. All samples were evaluated by Tuj1/NF200 and S100/MBP triple immunofluorescent staining. The primary antibodies were anti-Tuj1 (1:100, Abcam, USA), anti-NF200 (1:250, Abcam, USA), anti-S100 beta (1:500, Abcam, USA), and anti-MBP (1:200, Abcam, USA). The slides were observed under an immunofluorescence microscope (Leica, USA). The experiment was repeated for five times.
Qian Y., Zhao X., Han Q., Chen W., Li H, & Yuan W. (2018). An integrated multi-layer 3D-fabrication of PDA/RGD coated graphene loaded PCL nanoscaffold for peripheral nerve restoration. Nature Communications, 9, 323.
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5

Autophagy Assessment in Primary Neurons

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Transmission electron microscopy was performed to assess autophagy of the primary neurons 24 h after treatment with agomiR or antagomiR. Fixed cells were post-fixed in 2% OsO4, dehydrated in graded alcohol and flat embedded in Epon 812 (Electron Microscopy Sciences, Fort Washington, PA, USA). Ultra-thin sections (100 nm) were prepared, stained with uranyl acetate and lead citrate, and examined by electron microscopy (Hitachi H7500, Japan).
Zhang Y., Liu C., Wang J., Li Q., Ping H., Gao S, & Wang P. (2016). MiR-299-5p regulates apoptosis through autophagy in neurons and ameliorates cognitive capacity in APPswe/PS1dE9 mice. Scientific Reports, 6, 24566.
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6

Microscopic Evaluation of Pertussis Adherence

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Transmission electron microscopy was used to evaluate the adherence of B. pertussis to PHEA and HBE cells. Infected cells on air-liquid interface membranes were fixed serially with 2.5% glutaraldehyde and 1% osmium tetroxide and 1% uranyl acetate. Samples were dehydrated serially with increasing concentrations of ethanol (25%, 50%, 75%, 100%) before embedding with EPON812 (Electron microscopy sciences, Washington, PA) following standard procedures (30 (link)). Slice sections 0.5-mm-thick were created with a diamond knife and stained with toluidine blue for light microscopy examination before visualization under a transmission electron microscope (JEOL lOOC-ASID).
Samples to be evaluated by scanning electron microscope (SEM) were dehydrated, embedded, and mounted onto metallic stubs with double-sided carbon tape, sputter coated with a thin layer of metals (gold and palladium) and visualized. Evaluation of B. pertussis-infected cells by light microscopy (LM) required formalin fixation of millicell membranes containing cells followed by embedding in paraffin. Microtome sections from the paraffin block were stained with Giemsa, toluidine blue, or hematoxylin and eosin before visualization under the LM.
Guevara C., Zhang C., Gaddy J.A., Iqbal J., Guerra J., Greenberg D.P., Decker M.D., Carbonetti N., Starner T.D., McCray PB J.r., Mooi F.R, & Gómez-Duarte O.G. (2015). Highly Differentiated Human Airway Epithelial Cells: a Model to Study Host cell-parasite Interactions in Pertussis. Infectious diseases (London, England), 48(3), 177-188.
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7

Platelet Ultrastructure Analysis by TEM

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To analyse platelet ultrastructure, washed platelets were fixed with 2.5% glutaraldehyde (16,210, Electron Microscopy Sciences) in 50 mM cacodylate buffer (12,201, pH 7.2; AppliChem). After embedding in epon 812 (14,900, Electron Microscopy Sciences), ultrathin sections were generated and stained with 2% uranyl acetate (22,400, Electron Microscopy Sciences) and lead citrate (17,800, Electron Microscopy Sciences). Samples were visualized with an EM900 transmission electron microscope (Carl Zeiss).
Stritt S., Nurden P., Favier R., Favier M., Ferioli S., Gotru S.K., van Eeuwijk J.M., Schulze H., Nurden A.T., Lambert M.P., Turro E., Burger-Stritt S., Matsushita M., Mittermeier L., Ballerini P., Zierler S., Laffan M.A., Chubanov V., Gudermann T., Nieswandt B, & Braun A. (2016). Defects in TRPM7 channel function deregulate thrombopoiesis through altered cellular Mg2+ homeostasis and cytoskeletal architecture. Nature Communications, 7, 11097.
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8

Transmission Electron Microscopy Protocol

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Transmission EM was performed as described previously13 (link). In brief, cells were fixed with 2.5% glutaraldehyde (GA) in 50 mM sodium cacodylate buffer (CaCo), supplemented with 2% sucrose, 50 mM KCl, 2.6 mM MgCl2 and 2.6 mM CaCl2, for 30 min at room temperature. After five washes with 50 mM CaCo, samples were incubated with 2% OsO4 in 25 mM CaCo for 40 min on ice, washed three times with EM-grade water and incubated in 0.5% uranyl acetate in water overnight at 4 °C. Samples were rinsed three times with EM-grade water, dehydrated in a graded ethanol series (from 40 to 100%) at room temperature, embedded in Epon 812 (Electron Microscopy Sciences) and polymerized for 2 days at 60 °C. After polymerization, ultrathin sections of 70 nm were obtained by sectioning with an ultramicrotome Leica EM UC6 (Leica Microsystems) and mounted on a slot grid. Sections were counterstained using 3% uranyl acetate in 70% methanol for 5 min and lead citrate (Reynold’s) for 2 min and examined by using a JEOL JEM-1400 (JEOL) operating at 80 kV and equipped with a 4 K TemCam F416 (Tietz Video and Image Processing Systems GmbH).
Tabata K., Prasad V., Paul D., Lee J.Y., Pham M.T., Twu W.I., Neufeldt C.J., Cortese M., Cerikan B., Stahl Y., Joecks S., Tran C.S., Lüchtenborg C., V’kovski P., Hörmann K., Müller A.C., Zitzmann C., Haselmann U., Beneke J., Kaderali L., Erfle H., Thiel V., Lohmann V., Superti-Furga G., Brügger B, & Bartenschlager R. (2021). Convergent use of phosphatidic acid for hepatitis C virus and SARS-CoV-2 replication organelle formation. Nature Communications, 12, 7276.
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9

Platelet Ultrastructure Analysis by EM

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Washed platelets in a concentration of 3 × 105 platelets/μl were fixed with 2.5% glutaraldehyde (Electron Microscopy Science) in cacodylate buffer (pH 7.2, AppliChem). Epon 812 (Electron Microscopy Science) was used to embed platelets. After generation of ultrathin sections, platelets were stained with 2% uranyl acetate (Electron Microscopy Science) and lead citrate (Electron Microscopy Science). Sections were analyzed on a Zeiss EM900 electron microscope. Platinum replica electron microscopy of spread platelets was performed as previously described (28 (link)).
Baumann J., Sachs L., Otto O., Schoen I., Nestler P., Zaninetti C., Kenny M., Kranz R., von Eysmondt H., Rodriguez J., Schäffer T.E., Nagy Z., Greinacher A., Palankar R, & Bender M. (2022). Reduced platelet forces underlie impaired hemostasis in mouse models of MYH9-related disease. Science Advances, 8(20), eabn2627.
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10

Ultrastructural Visualization of PE N/MPL Uptake

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To assess the possible PE N/MPL uptake by vaginal cells and highlight the presence of intracellular PE N/MPLs, we performed TEM experiments. VK2 E6/E7 cells, cultured and treated as described above, were fixed in 2% glutaraldehyde (Electron Microscopy Sciences, Hatfield, PA, USA) in PBS for 24 h at 4 °C. After collection, cells were washed three times in PBS, post-fixed for 2 h in 1% OsO4 (Electron Microscopy Sciences), dehydrated in graded acetones and embedded in Epon-812 (Electron Microscopy Sciences) for 48 h at 60 °C. Ultrathin sections (60 nm) were cut with a Reichert ultramicrotome (Leica Microsystems, Wetzlar, Germany), mounted on copper grids, counterstained with uranyl-acetate replacement stain and lead citrate (Electron Microscopy Sciences) and finally examined with a Philips CM10 (TEM) and/or a Fei-Philips Morgagni 268D transmission electron microscope (FEI, Eindhoven, The Netherlands). Furthermore, to mimic the formation of a protein corona [9 (link)], we employed PE N/MPLs coated with keratin from human epidermis (Sigma-Aldrich), by incubation for 24 h at 4 °C prior to the treatment.
Pontecorvi P., Ceccarelli S., Cece F., Camero S., Lotti L.V., Niccolai E., Nannini G., Gerini G., Anastasiadou E., Scialis E.S., Romano E., Venneri M.A., Amedei A., Angeloni A., Megiorni F, & Marchese C. (2023). Assessing the Impact of Polyethylene Nano/Microplastic Exposure on Human Vaginal Keratinocytes. International Journal of Molecular Sciences, 24(14), 11379.
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