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Ultracut e microtome

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The Ultracut E microtome is a precision instrument designed for the preparation of ultra-thin sections of materials for examination under an electron microscope. It features a sturdy construction and advanced mechanics to ensure accurate and reliable sectioning of samples.

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Lab products found in correlation

Lx112, by Polysciences (2 mentions) Diamond histoknife, by Electron Microscopy Sciences (2 mentions) Slice view g2 operating software, by Thermo Fisher Scientific (1 mentions) Quanta 3d feg, by Thermo Fisher Scientific (1 mentions) Diamond knife, by Electron Microscopy Sciences (1 mentions) Glutaraldehyde, by Agar Scientific (1 mentions) Cm12 tem, by Philips (1 mentions) Talos l120c g2 transmission electron microscope, by Thermo Fisher Scientific (1 mentions) H 7700, by Hitachi (1 mentions) Toluidine blue, by Merck Group (1 mentions) Paraformaldehyde (pfa), by Merck Group (1 mentions) Glutaraldehyde, by Merck Group (1 mentions) H 7100 transmission electron microscope, by Hitachi (1 mentions) Embed 812, by Electron Microscopy Sciences (1 mentions)

16 protocols using ultracut e microtome

1

Ultrastructural Imaging of Dentate Gyrus Neurons

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The embedded slices were placed on a metal stub and further trimmed with glass and diamond knives in an ultramicrotome (Ultracut E microtome, Leica, Wetzlar, Germany). The slices were coated with a protective layer of carbon, which prevented any charge. The metal stub with the slices was set on the stage of FIB/SEM. The serial section images in the middle molecular layer of the dorsal DG were automatically obtained by FIB/SEM (Quanta 3D FEG, FEI, Hillsboro, OR, USA). Serial images of the block face were acquired by repeated cycles of sample surface milling and imaging using the Slice & View G2 operating software (FEI). The milling was performed with a gallium ion beam at 30 kV with a current of 1.0 nA or 3.0 nA (Golgi stain). The milling pitch was set to 15 or 30 (Golgi stain) nm/step. The images were acquired at an accelerating voltage of 2.5 kV. The other acquisition parameters were as follows: dwell time = 6 s/pixel, pixel size = 4.9 and 14.6 (Golgi stain) nm/pixel.
Kitahara Y., Ohta K., Hasuo H., Shuto T., Kuroiwa M., Sotogaku N., Togo A., Nakamura K.I, & Nishi A. (2016). Chronic Fluoxetine Induces the Enlargement of Perforant Path-Granule Cell Synapses in the Mouse Dentate Gyrus. PLoS ONE, 11(1), e0147307.
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2

Morphology Analysis of TPE Nanocomposites

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The morphology of TPE and TPE nanocomposites was studied using transmission electron microscopy (TEM) (Philips model STEM CM12) with an accelerating voltage of 100 kV. An ultrathin piece of sample was prepared using a Leica ultracut E microtome with a cryo FC4E attachment at −100 °C.
Tarawneh M.A., Saraireh S.A., Chen R.S., Ahmad S.H., Al- Tarawni M.A, & Yu L.J. (2020). Gamma irradiation influence on mechanical, thermal and conductivity properties of hybrid carbon nanotubes/montmorillonite nanocomposites. Radiation Physics and Chemistry, 179, 109168.
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3

Ultrastructural Analysis of Nerve Repair

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After excision and dissection of the middle of the repair constructs, samples that had been fixed in 4% (w/v) paraformaldehyde in PBS for 24 h were transferred to 3% glutaraldehyde (Agar Scientific) in 0.1 M cacodylate buffer. These were post-fixed in 1% (w/v) osmium tetroxide in PBS, dehydrated through a graded series of ethanol incubations, flat-embedded in TAAB embedding resin and polymerized at 60 °C for 48 h. Semi-thin sections of 0.5 μm were cut using a diamond knife on a Ultracut E microtome (Leica, UK), dried onto microscope slides and stained with 1% (w/v) toluidine blue with added 5% (w/v) sodium borate. Ultrathin sections of 70 nm were cut with a diamond knife (Diatome, UK) and collected on copper slot grids with Formvar/carbon support films. Sections were counter-stained with ethanol based uranyl acetate and Reynolds’ lead citrate before examination in a Philips CM12 TEM. Ultrathin sections were imaged at a column magnification of × 2000 from the five areas of greatest tissue density as identified from the respective stained semi-thin sections. Image J software was used to measure axon and fibre diameter, from which myelin thickness and G-ratio were calculated.
O’Rourke C., Day A.G., Murray-Dunning C., Thanabalasundaram L., Cowan J., Stevanato L., Grace N., Cameron G., Drake R.A., Sinden J, & Phillips J.B. (2018). An allogeneic ‘off the shelf’ therapeutic strategy for peripheral nerve tissue engineering using clinical grade human neural stem cells. Scientific Reports, 8, 2951.
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4

Ultrastructural Analysis of Parasitic Egress

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Parasites were pretreated for 72 h (pi3ki parasites) or 48 h (pikfyvei parasites) with or without 1.5 μg/ml ATc, collected promptly after egress and inoculated onto new HFF monolayers in presence of ATc during 24 hours. Infected HFF monolayers on coverslips were fixed for 4 hours at room temperature with 2.5% glutaraldehyde (EMS) in 0.1M phosphate buffer pH7.2, washed in buffer and post-fixed for 1 hour in 1% OsO4, washed in water and stained overnight in 2% uranylacetate. Coverslips were then dehydrated in ethanol series and embedded in Epon (Embed 812, EMS). Ultrathin sections were prepared with a Leica ultracut E microtome, contrasted with 2% uranylacetate in ethanol and lead citrate and observed with a JEOL 1200E electron microscope.
Daher W., Morlon-Guyot J., Sheiner L., Lentini G., Berry L., Tawk L., Dubremetz J.F., Wengelnik K., Striepen B, & Lebrun M. (2014). Lipid kinases are essential for apicoplast homeostasis in Toxoplasma gondii. Cellular microbiology, 17(4), 559-578.
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5

Ultrastructural Analysis of Synaptosomes

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For ultrastructural analysis, purified synaptosomes were fixed in 2.5% glutaraldehyde in 0.1M cacodylate buffer, pH 7.2, post-fixed in 1% osmium tetroxide in cacodylate buffer 0.1 M, pH 7.2, en bloc stained with a 1% aqueous solution of uranyl acetate. The dehydration was performed through a graded ethanol series. Samples were then embedded in LX112 (Polysciences Inc., Warrington, PA, USA), polymerized for 12 h at 42 °C, followed by 48 h at 60 °C. A Leica Ultracut E microtome was used to prepare grey-silver ultrathin sections that were then stained with uranyl acetate and lead citrate. All images were acquired using a FEI Talos L120C G2 Transmission Electron Microscope (Thermo Scientific™, Monza, Italy).
Cervetto C., Amaroli A., Amato S., Gatta E., Diaspro A., Maura G., Signore A., Benedicenti S, & Marcoli M. (2023). Photons Induce Vesicular Exocytotic Release of Glutamate in a Power-Dependent Way. International Journal of Molecular Sciences, 24(13), 10977.
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6

Ultrastructural Changes in Renal Mitochondria

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Transmission electron microscopy was used to investigate the ultrastructural changes in mitochondria. After 0 h, 24 h or 48 h of hypothermic preservation, the renal tissue was diced into 1 mm3 pieces and further fixed using 2.5% glutaric dialdehyde for 4 h. The pieces were postfixed in 2% osmium tetroxide for 30 min, dehydrated in graded alcohol, transferred to propylene oxide, and gradually embedded in blocks of Epon 812 resin for 2 days at 60 °C. Eighty-nanometer sections were collected using a diamond histoknife (Diatome) on an Ultracut E microtome (Leica) and then mounted on a copper mesh and stained with uranyl acetate and lead nitrate. The ultrastructural changes in mitochondria was identified by a transmission electron microscope (Hitachi H-7700).
Xu L., Xu Y., Zhu Z., Gu H., Chen C, & Chen J. (2021). Tanshinone IIA attenuates renal injury during hypothermic preservation via the MEK/ERK1/2/GSK-3β pathway. BMC Complementary Medicine and Therapies, 21, 257.
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7

Quantifying Glomerular Basement Membrane Thickness

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Sections from the renal cortex area of the right kidneys were diced into 1–2 mm pieces and fixed in 4% PFA for 4 h followed by post-fixation with 1% osmium tetroxide for 30 min. Then, sections were then dehydrated in graded alcohol, transferred to propylene oxide and embedded in Epon 812 resin in blocks. Eighty nanometer sections were collected using a diamond histoknife (Diatome) on an Ultracut E microtome (Leica), and then mounted on a copper mesh and stained with uranyl acetate and lead nitrate. The UTHSCSA Image Tools 3.0 (University of Texas Medical School at San Antonio) was used to determine the thickness of the glomerular basement membrane. Glomerular basement membrane (GBM) thickness was determined by the orthogonal intercept method.19 (link)
Xu S., He L., Ding K., Zhang L., Xu X., Wang S, & Qian X. (2020). Tanshinone IIA Ameliorates Streptozotocin-Induced Diabetic Nephropathy, Partly by Attenuating PERK Pathway-Induced Fibrosis. Drug Design, Development and Therapy, 14, 5773-5782.
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8

Transmission Electron Microscopy of Cellular Precipitates

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The precipitate obtained from differential centrifugation and filtration was used for transmission electron microscopy. The supernatant was removed and kept at 4°C overnight. The precipitate was fixed with PBS buffer containing 2% paraformaldehyde (Sigma-Aldrich) and 2% glutaraldehyde (Sigma-Aldrich) and placed at room temperature for 10 min. Then, the pellet was washed with PBS buffer and fixed with 1% OsO4 (Taab; Aldermaston, Berks, UK) for 1 hour. Finally, the samples were dehydrated in a series of graded ethanol and embedded in epoxy resin (Epon 812). Semithin and ultrathin sections were cut on an Ultracut E microtome (Leica, Nussloch, Germany). The semithin sections (1 mm) were stained with toluidine blue (Sigma-Aldrich) and examined under an optical microscope. Ultrathin sections (60-80 nm) were stained with 2% uranyl acetate and lead citrate, and the samples were examined under an FEI Morgagni 268(D) transmission electron microscope (FEI NanoPorts, Hillsboro, USA).
Tian J., Wang Y., Ding M., Zhang Y., Chi J., Wang T., Jiao B., Jian Z., Yi X., Huang Y., Liu L., Li K., Chen J., Wang G., Gao T., Li C, & Li Q. (2021). The Formation of Melanocyte Apoptotic Bodies in Vitiligo and the Relocation of Vitiligo Autoantigens under Oxidative Stress. Oxidative Medicine and Cellular Longevity, 2021, 7617839.
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9

Histological Analysis of Mice Spinal Cord and Tissues

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Mice were deeply anaesthetized using an overdose of pentobarbital (200 mg/ml) containing lidocaine (20 mg/ml) and perfused through the left ventricle with cold 4% paraformaldehyde (PFA) in phosphate-buffered saline (PBS).
Toluidine blue (TB) staining and electron microscopy (EM) analyses: For EM and TB analyses of axons and myelin, 1 mm segments of the thoracic spinal cords were processed as previously described [9 ]. One µm thick sections were obtained with a Leica Ultracut E microtome and stained with 1% TB solution.
Paraffin histopathology and immunohistochemical analysis: Spinal cords were removed and tissue segments containing the lesion area were paraffin-embedded and cut into 10 parallel series of 15µm thick microtome sections. Furthermore, in naïve mice lung, heart, liver, diaphragm, spleen, the anterior tibialis muscle and small intestine biopsies were imbedded in paraffin and cut into 5µm thick sections. Sections were then stained using Hematoxylin and Eosin (H&E) [3 (link)]. For stereological analysis of the total number of astrocytes in the hilus of the hippocampus, brains were processed as previously described [18 (link)] and cut horizontally into six 60µm-thick parallel series of free floating vibratome sections and cryo-protected in de Olmo’s solution.
Ellman D.G., Novrup H.G., Jørgensen L.H., Lund M.C., Yli-Karjanmaa M., Madsen P.M., Vienhues J.H., Dursun S., Bethea J.R., Lykke-Hartmann K., Brambilla R, & Lambertsen K.L. (2017). Neuronal Ablation of IKK2 Decreases Lesion Size and Improves Functional Outcome after Spinal Cord Injury in Mice. JSM neurosurgery and spine, 5(3), 1090.
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10

Estimating Basal Membrane Thickness

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The method of estimating the thickening of the basal membrane was described previously (14) . In brief, specimens were fixed with Karnovsky fixative, dehydrated with an alcohol series, and embedded in araldite (SERVA). Sections of 70-nm thickness were cut using an Ultracut E microtome (Leica). Sections were analyzed with a Zeiss EM 900 or 910 transmission electron microscope (Zeiss, Oberkochen, Germany) and macrographs were taken with a CCDK2 camera (TRS-Tröndle, Dünzelbach, Germany). Basal membrane thickness was estimated with ImageSP software (TRS-Tröndle).
Chen J., Fleming T., Katz S., Dewenter M., Hofmann K., Saadatmand A., Kronlage M., Werner M.P., Pokrandt B., Schreiter F., Lin J., Katz D., Morgenstern J., Elwakiel A., Sinn P., Gröne H.J., Hammes H.P., Nawroth P.P., Isermann B., Sticht C., Brügger B., Katus H.A., Hagenmueller M, & Backs J. (2021). CaM Kinase II-δ Is Required for Diabetic Hyperglycemia and Retinopathy but Not Nephropathy. Diabetes, 70(2).
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