Poly bed 812 epoxide resin

Manufactured by Polysciences

Sourced in United Kingdom

Poly/Bed 812 is an epoxide resin. It is a two-part resin system that is used for embedding and encapsulating samples for various applications, including electron microscopy and materials science. The resin cures at room temperature and produces a hard, durable final product.

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Lab products found in correlation

Bx51 microscope, by Jenoptik (1 mentions) H 7650, by Hitachi (1 mentions) Tecnai 120kv transmission electron microscope, by Thermo Fisher Scientific (1 mentions) Jem 100s, by JEOL (1 mentions)

4 protocols using poly bed 812 epoxide resin

Microscopy analysis of live and fixed cells

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Differential interference contrast micrographs of live and formaldehyde-fixed cells were taken on an Olympus BX51 microscope equipped with a ProgRes CFscan CCD camera (Jenoptik, Jena, Germany) using a 60×/1.40 oil-immersion objective lens.
For thin-section electron microscopy, log-phase cells in TAP medium were fixed by addition of an equal volume of TAP containing 5% glutaraldehyde (EM grade; EM Sciences, Hatfield, PA). After 15 min, cells were harvested by gentle centrifugation and resuspended in 2.5% glutaraldehyde in 0.1 M Na cacodylate (pH 7.4) for 45 min. Cells were then washed five times with cacodylate buffer and postfixed with 1% OsO₄ and 0.8% K₃Fe(CN)₆ in cacodylate buffer for 60 min. Following multiple washes with distilled water, samples were stained en bloc with 1% aqueous uranyl acetate, dehydrated through an ethanol series, transitioned to propylene oxide, and embedded in Poly/Bed 812 epoxide resin (Polysciences, Warrington, PA). Ultrathin sections with a nominal thickness of 55 nm were picked up on unsupported 300-mesh copper grids, poststained with 6.25% uranyl acetate in 50% methanol, and examined in a Hitachi H-7650 transmission electron microscope operating at 80 kV.
All micrographs were cropped and adjusted for brightness/contrast using Adobe Photoshop CS4.

Patel-King R.S., Sakato-Antoku M., Yankova M, & King S.M. (2019). WDR92 is required for axonemal dynein heavy chain stability in cytoplasm. Molecular Biology of the Cell, 30(15), 1834-1845.

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Glutaraldehyde-Osmium Fixation of Cells

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Cells in R medium were fixed by adding an equal volume of R medium containing 5% glutaraldehyde (EM grade; EM Sciences, Hatfield, PA). After 15 min, cells were pelleted by low-speed centrifugation and resuspended in 2.5% glutaraldehyde in 0.1 M Na cacodylate buffer at pH 7.4 for 45 mins. Following multiple washes with cacodylate buffer, samples were postfixed with 1% OsO₄ and 0.8% K₄Fe(CN)₆ in cacodylate buffer. Note that this step differs from descriptions in our previous studies as the ferrocyanide, rather than the ferricyanide (K₃Fe(CN)₆), salt was used. After several washes with water, cells were stained en bloc with 1% uranyl acetate (wt/vol; aq.), dehydrated through an ethanol series, transitioned into propylene oxide, and finally embedded in Poly/Bed 812 epoxide resin (Polyscience, Warrington, PA). Ultrathin sections of ∼55 nm were placed on 300 mesh copper grids and stained with 6.25% uranyl acetate in 50% methanol. Cells were imaged in a Hitachi H-7650 transmission electron microscope operating at 80 kV. Micrographs were cropped and adjusted for brightness/contrast using Adobe Photoshop ver. 22.4.2.

Sakato-Antoku M, & King S.M. (2023). Outer-arm dynein light chain LC1 is required for normal motor assembly kinetics, ciliary stability, and motility. Molecular Biology of the Cell, 34(7), ar75.

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Transmission Electron Microscopy Tissue Processing

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The tissue samples were collected and fixed in 3% glutaraldehyde.
Processing was done with a Leica EM TP processor (Leica Microsystems Inc., North
Deerfield, IL). Samples were rinsed with buffer, post-fixed in 2% osmium
tetroxide in phosphate buffer, rinsed in distilled water and dehydrated in an
ethanol series transitioning into propylene oxide. The samples were then
infiltrated with Poly/Bed 812 epoxide resin (Polysciences). After
polymerization, selected blocks were trimmed and semithin sections
(approximately 0.5 μm thick) were cut, mounted on glass slides, stained
with 1% toluidine blue in 1% sodium borate, and examined with a light microscope
to ascertain areas of interest. After trimming block faces down to areas of
interest, ultrathin sections (70–90 nm thick) were subsequently cut,
placed on 100 mesh formvar copper grids and stained with uranyl acetate and lead
citrate. Digital images were captured with an Olympus Mega View III side mount
camera attached to an FEI Tecnai 120KV transmission electron microscope.

Arao Y., Stumpo D.J., Hoenerhoff M.J., Tighe R.M., Yu Y.R., Sutton D., Kashyap A., Beerman I, & Blackshear P.J. (2023). Lethal eosinophilic crystalline pneumonia in mice expressing a stabilized Csf2 mRNA. FASEB journal : official publication of the Federation of American Societies for Experimental Biology, 37(8), e23100.

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Leaf Ultrastructure Microscopy Protocol

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Specimens for LM and TEM analysis were obtained from the central part of healthy green leaf blades (second or third leaf) and were fixed in 3.5% glutaraldehyde solution in phosphate buffer with the pH of 7.0 for 12 h at room temperature, followed by secondary fixation in 2.5% osmium tetroxide solution. The specimens were rinsed and dehydrated in a graded series of alcohols and acetone, and were embedded in Poly Bed 812 epoxide resin (Polyscience). Microtome sections were prepared in the Leica Ultracut R microtome using Diatome diamond knives. Semi-thin sections, 1.5 μm thick, were placed on slides and stained with toluidine blue and azure B. The specimens were mounted with a drop of glycerin. They were analyzed under the Nikon Eclipse 80i light microscope with compatible hardware and software (NIS ELEMENTS) for digital image recording. Ultra-thin sections, 60-90 nm thick, were mounted on nickel grids with 300 mesh squares. Immediately before examination, saturated aqueous uranyl acetate solution and lead citrate were added to impart contrast to the specimens. The specimens were examined and electronograms were obtained simultaneously under two transmission electron microscopes-JEOL JEM 100S and JEOL 1400. JEOL JEM 100S supports analog image recording, whereas JEOL 1400 is equipped with hardware and iTEM software for recording data files.

GieÅwanowska I., GÃ³recki Ryszard J..., GÃ³rniak D.o.r.o.t.a., Kellmann-SopyÅa W.i.o.l.e.t.a, & Pastorczyk M.a.r.t.a. (2015). Morphological and Ultrastructural Changes of Organelles in Leaf Mesophyll Cells of the Arctic and Antarctic Plants of Poaceae Family Under Cold Influence. Arctic, antarctic, and alpine research., 47(1).

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