Ultracut s microtome

Manufactured by JEOL

Sourced in United Kingdom

The Ultracut-S microtome is a laboratory instrument designed for the precise cutting of ultra-thin sections, typically used in electron microscopy sample preparation. It features precise control and high-quality sample sectioning capabilities.

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Lab products found in correlation

1200 ex transmission electron microscope, by JEOL (14 mentions) Amt 2k ccd camera, by JEOL (6 mentions) Tecnaig spirit biotwin, by JEOL (3 mentions) Tecnaig2 spirit biotwin, by JEOL (2 mentions) Jsm 7500f sem instrument, by JEOL (1 mentions) Sp5 x mp inverted confocal microscope, by Leica (1 mentions) 1200 ex transmission, by JEOL (1 mentions) Jem 1400, by JEOL (1 mentions) Spurr s low viscosity resin, by Electron Microscopy Sciences (1 mentions) 1200ex electron microscope, by JEOL (1 mentions) Karnovsky s fixative, by Electron Microscopy Sciences (1 mentions) Cx41 microscope, by Olympus (1 mentions) Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (1 mentions) Cm120 electron microscope, by Philips (1 mentions)

48 protocols using ultracut s microtome

Electron Microscopy Sample Preparation

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A pellet of cells was fixed for at least 2 h at RT in fixative (2.5% Glutaraldehyde 1.25% Paraformaldehyde and 0.03% picric acid in 0.1 M sodium cacodylate buffer (pH 7.4)), washed in 0.1 M cacodylate buffer and post-fixed with 1% Osmiumtetroxide (OsO4)/1.5% Potassiumferrocyanide (KFeCN6) for 1 h, washed 2x in water, 1x Maleate buffer (MB) 1x and incubated in 1% uranyl acetate in MB for 1 h followed by 2 washes in water and subsequent dehydration in grades of alcohol (10 mins each; 50%, 70%, 90%, 2X10 mins 100%). The samples were then put in propyleneoxide for 1 h and infiltrated overnight in a 1:1 mixture of propyleneoxide and TAAB (TAAB Laboratories Equipment Ltd, https://taab.co.uk). The following day the samples were embedded in TAAB Epon and polymerized at 60°C for 48 h. Ultrathin sections (about 60 nm) were cut on a Reichert Ultracut-S microtome, picked up on to copper grids stained with lead citrate and examined in a JEOL 1200EX Transmission electron microscope or a TecnaiG² Spirit BioTWIN and images were recorded with an AMT 2k CCD camera.

Tung Yiu S.P., Guo R., Zerbe C., Weekes M.P, & Gewurz B.E. (2022). Epstein-Barr virus BNRF1 destabilizes SMC5/6 cohesin complexes to evade its restriction of replication compartments. Cell reports, 38(10), 110411.

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Multimodal Imaging Characterization of Nanoparticles

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3D-reconstructed fluorescence microscopy imaging was performed using a Leica SP5 X MP inverted confocal microscope equipped with a 60 × 1.42 NA oil immersion objective, with a set of standard filters for DAPI/CFP/FITC/AF488/AF568/Cy5/AF647. Image processing and 3D models were analyzed and generated with Imaris (Bitplane) software using the maximum intensity projection. UV-Vis absorption and fluorescence measurements were conducted on an Infinite M Nano microplate reader (Tecan Group, Switzerland). Transmission electron microscopy (TEM) was performed on a Tecnai G2 F20 S-TWIN TEM instrument, operating at a voltage of 100 kV (FEI USA, Inc.). Scanning electron microscopy (SEM) was performed on a JEOL JSM-7500F SEM instrument. Particle zeta potential was measured by dynamic light scattering (DLS) on Zetasizer Nano ZSP (Malvern, UK). Ultrathin sections (about 80 nm) were cut on a Reichert Ultracut-S microtome, picked up onto copper grids stained with lead citrate, and examined in a JEOL 1200EX Transmission electron microscope, and images were recorded with an AMT 2k CCD camera.

Pan J., Gong G., Wang Q., Shang J., He Y., Catania C., Birnbaum D., Li Y., Jia Z., Zhang Y., Joshi N.S, & Guo J. (2022). A single-cell nanocoating of probiotics for enhanced amelioration of antibiotic-associated diarrhea. Nature Communications, 13, 2117.

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Transmission Electron Microscopy of Tumor Samples

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Small tumor fragments (1–2 mm³) were fixed in FGP (2.5% glutaraldehyde, 1.25% paraformaldehyde and 0.03% picric acid in 0.1 M sodium cacodylate buffer, pH 7.4) at 4 ˚C. Three tumor samples from each group (drug or control) were analyzed at the Electron Microscopy Core Facility (Harvard Medical School, Boston).
Tissue samples were postfixed with 1% osmium tetroxide (OsO4)/1.5% potassium ferrocyanide (KFeCN6) for 1 hour, washed in water 2 times, 1 time in 50mM maleate buffer (pH 5.15, MB) and incubated in 1% uranyl acetate in MB for 1hr followed by 1 wash in MB, 2 washes in water and subsequent dehydration in grades of alcohol (10min each; 50%, 70%, 90%, 2×10min 100%). The samples were left in propyleneoxide for 1 hr and infiltrated overnight in a 1:1 mixture of propyleneoxide and TAAB Epon (TAAB Laboratories Equipment Ltd, https://taab.co.uk). The following day the samples were embedded in TAAB Epon and polymerized at 60ºC for 48 hr. Ultrathin sections (about 80nm) were cut on a Reichert Ultracut-S microtome, picked up on to copper grids stained with lead citrate, and examined in a JEOL 1200EX Transmission electron microscope or a TecnaiG² Spirit BioTWIN. Images were recorded with an AMT 2k CCD camera.

Feng Y., Mischler W.J., Gurung A.C., Kavanagh T.R., Androsov G., Sadow P.M., Herbert Z.T, & Priolo C. (2020). Therapeutic targeting of the secreted lysophospholipase D autotaxin suppresses tuberous sclerosis complex-associated tumorigenesis. Cancer research, 80(13), 2751-2763.

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Ultrastructural Analysis of Lgr5-lacZ Colon

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Two Lgr5-lacZ male mice, 6–8 weeks old, were used per time point. Colonic samples were fixed in 2.5% glutaraldehyde plus 2% paraformaldehyde in 0.075 M sodium cacodylate buffer for 1 hour at 4°C and postfixed in 2% osmium tetroxide in double-distilled water. Samples were then rinsed in double-distilled water and dehydrated in a graded alcohol series of 50%, 75%, and 95% through absolute alcohol, followed by propylene oxide, with overnight incubation in 1:1 propylene oxide/poly bed 812 epoxy resin (manufacturer unknown). Ultrathin sections, obtained with a Reichert Ultracut S microtome, were stained with uranyl acetate and lead citrate and photographed using a Jeol 1200 EX transmission electron microscope. Experiments were performed in the Electron Microscopy Facility at the Sloan Kettering Institute.

Klingler S., Hsu K.S., Hua G., Martin M.L., Adileh M., Baslan T., Zhang Z., Paty P.B., Fuks Z., Brown A.M, & Kolesnick R. (2022). Disruption of the crypt niche promotes outgrowth of mutated colorectal tumor stem cells. JCI Insight, 7(5), e153793.

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Zygote Ultrastructural Analysis Protocol

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Sample preparation was performed by following the protocol of Harvard Medical School EM facility. Briefly, zygotes were fixed overnight at 4°C in 2.5% Glutaraldehyde, 1.25% Paraformaldehyde and 0.03% picric acid in 0.1 M sodium cacodylate buffer (pH 7.4). Samples were washed in 0.1 M cacodylate buffer, fixed with 1% Osmiumtetroxide (OsO₄)/1.5% Potassiumferrocyanide (KFeCN₆) for 2 h, washed in water 3 times and incubated in 1% aqueous uranyl acetate for 1 h, followed by 2 washes in water and subsequent dehydration in grades of alcohol (10 min each; 50%, 70%, 90%, 2×10 min 100%). The samples were then put in propyleneoxide for 1 h and infiltrated overnight in a 1:1 mixture of propyleneoxide and Spurr’s resin (EMS, Hatfield, PA). On the following day, the samples were embedded in Spurr’s and polymerized at 60°C for 48 h.
Ultra-thin sections (about 60 nm) were cut on a Reichert Ultracut-S microtome, picked up on to copper grids, stained with lead citrate and examined in a JEOL 1200EX Transmission electron microscope. Images were recorded with an AMT 2k CCD camera.

Inoue A, & Zhang Y. (2014). Nucleosome assembly is required for nuclear pore complex assembly in mouse zygotes. Nature structural & molecular biology, 21(7), 609-616.

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Bacterial TEM Sample Preparation Protocol

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To prepare exponential-phase bacterial TEM samples, an overnight culture of USA400 MW2 (prepared in TSB) was diluted 1:300 in TSB and grown at 37 °C with 200 rpm shaking until an OD₆₀₀ of 0.6 was reached. Compound treatments were performed (4 μM V or V–r8) for 90 min. To prepare stationary-phase bacterial samples, an overnight culture of USA400 MW2 was pelleted and washed 1× in PBS and then diluted to an OD₆₀₀ of 1.0 in PBS. Compound treatments were performed at 8 μM for V and V–r8 for 90 min. Upon treatment, all cells were then pelleted at 10000g at 4 °C for 10 min and then resuspended in fixative (2% glutaraldehyde and 4% paraformaldehyde in 0.1 M sodium cacodylate, pH 7.4). Upon three PBS washes to remove excess fixative, the samples were enrobed in gelatin, placed in a cold block, and postfixed with 1% OsO₄ in phosphate buffer. Samples were then washed three times with ddH₂O, stained with 1% uranyl acetate overnight at 4 °C, and then subsequently dehydrated with 95% EtOH and embedded in Epon. A Leica Ultracut S microtome was used to prepare thin sections, which were viewed on a JEOL JEM-1400.

Antonoplis A., Zang X., Huttner M.A., Chong K.K., Lee Y.B., Co J.Y., Amieva M.R., Kline K.A., Wender P.A, & Cegelski L. (2018). A Dual-Function Antibiotic-Transporter Conjugate Exhibits Superior Activity in Sterilizing MRSA Biofilms and Killing Persister Cells. Journal of the American Chemical Society, 140(47), 16140-16151.

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Ultrastructural Analysis of Mouse Left Ventricular Tissue

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Mice were sacrificed and left ventricular tissue was isolated under a dissecting microscope. Left ventricular tissue was cut into small pieces (1-2mm cubes) and perfused in 1% glutaraldehyde/3% paraformaldehyde in cardioplegic buffer (5% dextrose, 30mmol/L KCl in PBS).^{30 (link)} After fixation the tissue was washed in 0.1M cacodylate buffer and postfixed with 1% Osmiumtetroxide (OsO4)/1.5% Potassium ferrocyanide (KFeCN6) for 1 hour, washed in water 3x and incubated in 1% aqueous uranyl acetate for 1hr followed by 2 washes in water and subsequent dehydration in grades of alcohol (10min each; 50%, 70%, 90%, 2x10min 100%). The samples were then put in propyleneoxide for 1 hr and infiltrated ON in a 1:1 mixture of propyleneoxide and TAAB Epon (Marivac Canada Inc. St. Laurent, Canada). The following day the samples were embedded in TAAB Epon and polymerized at 60°C for 48 hrs. Ultrathin sections (about 80nm) were cut on a Reichert Ultracut-S microtome, picked up on to copper grids, stained with lead citrate and examined in a JEOL 1200EX Transmission electron microscope or a TecnaiG² Spirit BioTWIN. Images were recorded with an AMT 2k CCD camera and saved as TIFF files.

Agarwal R., Wakimoto H., Paulo J.A., Zhang Q., Reichart D., Toepfer C., Sharma A., Tai A.C., Lun M., Gorham J., DePalma S.R., Gygi S.P., Seidman J.G, & Seidman C.E. (2022). Pathogenesis of Cardiomyopathy Caused by Variants in ALPK3, an Essential Pseudokinase in the Cardiomyocyte Nucleus and Sarcomere. Circulation, 146(22), 1674-1693.

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Comprehensive Electron Microscopy Protocol

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Samples were fixed in the routine fixative [2.5% Glutaraldehyde 1.25% Paraformaldehyde and 0.03% picric acid in 0.1 M sodium cacodylate buffer (pH 7.4)] for at least 2 h at room temperature, washed in 0.1M cacodylate buffer and postfixed with 1% Osmiumtetroxide (OsO4)/1.5% Potassiumferrocyanide (KFeCN6) for 1 h, washed 2 × in water, 1 × Maleate buffer (MB) 1x and incubated in 1% uranyl acetate in MB for 1hr followed by 2 washes in water and subsequent dehydration in grades of alcohol (10 min each; 50%, 70%, 90%, 2 × 10 min 100%). The samples were then put in propyleneoxide for 30 min and infiltrated ON in a 1:1 mixture of propyleneoxide and TAAB Epon (Marivac Canada Inc. St. Laurent, Canada). The following day the samples were embedded in TAAB Epon and polymerized at 60°°C for 48 h. Ultrathin sections (about 60 nm) were cut on a Reichert Ultracut-S microtome, picked up on to copper grids stained with lead citrate and examined in a JEOL 1200EX Transmission electron microscope and images were recorded with an AMT 2k CCD camera.

Xu C., Ericsson M, & Perrimon N. (2018). Understanding cellular signaling and systems biology with precision: A perspective from ultrastructure and organelle studies in the Drosophila midgut. Current opinion in systems biology, 11, 24-31.

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Ultrastructural Analysis of Biological Tissues

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For transmission electron microscopy analysis, tissues were fixed in 2.5% Glutaraldehyde 1.25% Paraformaldehyde and 0.03% picric acid in 0.1 M sodium cacodylate buffer (pH 7.4). Small tissue pieces (1–2mm cubes) were fixed for at least 2 hours at RT in the above fixative, washed in 0.1M cacodylate buffer and postfixed with 1% Osmium tetroxide (OsO4)/1.5% Potassium ferrocyanide (KFeCN6) for 1 hour, washed in water 3x and incubated in 1% aqueous uranyl acetate for 1hr followed by 2 washes in water and subsequent dehydration in grades of alcohol (10min each; 50%, 70%, 90%, 20×10min 100%). The samples were then put in propyleneoxide for 1 hr and infiltrated ON in a 1:1 mixture of propyleneoxide and TAAB Epon (Marivac Canada Inc. St. Laurent, Canada). The following day the samples were embedded in TAAB Epon and polymerized at 60°c for 48 hrs. Ultrathin sections (~60nm) were cut on a Reichert Ultracut-S microtome, picked up on to copper grids stained with lead citrate and examined in a JEOL 1200EX Transmission electron microscope or a TecnaiG² Spirit BioTWIN and images were recorded with an AMT 2k CCD camera.

Yissachar N., Zhou Y., Ung L., Lai N.Y., Mohan J.F., Ehrlicher A., Weitz D.A., Kasper D.L., Chiu I.M., Mathis D, & Benoist C. (2017). An intestinal organ culture system uncovers a role for the nervous system in microbe-immune crosstalk. Cell, 168(6), 1135-1148.e12.

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Ultrastructural Cellular Imaging via EM

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Electron microscopy imaging was performed in the Harvard Medical School Electron Microscopy Facility. Fixative solution containing 2.5% glutaraldehyde, 1.25% paraformaldehyde, 0.03% picric acid in 0.1 M sodium cacodylate buffer, pH 7.4, was added in a 1:1 ratio to cells grown to 70% confluency for 1 h at room temperature. The cells were then postfixed for 30 min in 1% osmium tetroxide (OsO₄)/1.5% potassium ferrocyanide (KFeCN₆), washed in water 3x, and incubated in 1% aqueous uranyl acetate for 30 min, followed by two washes in water and subsequent dehydration in grades of alcohol (5 min each; 50, 70, 95%, 2 × 100%). Cells were removed from the dish in propyleneoxide, pelleted at 500×g for 3 min, and infiltrated for 2 h to overnight in a 1:1 mixture of propyleneoxide and TAAB Epon (Marivac Canada Inc., St. Laurent, Canada). The samples subsequently embedded in TAAB Epon and polymerized at 60 °C for 48 h. Ultrathin sections (~60 nm) were cut on a Reichert Ultracut-S microtome, picked up on to copper grids stained with lead citrate and examined in a JEOL 1200EX Transmission electron microscope or a TecnaiG² Spirit BioTWIN and images were recorded with an AMT 2k CCD camera.

Boland S., Swarup S., Ambaw Y.A., Malia P.C., Richards R.C., Fischer A.W., Singh S., Aggarwal G., Spina S., Nana A.L., Grinberg L.T., Seeley W.W., Surma M.A., Klose C., Paulo J.A., Nguyen A.D., Harper J.W., Walther T.C, & Farese RV J.r. (2022). Deficiency of the frontotemporal dementia gene GRN results in gangliosidosis. Nature Communications, 13, 5924.

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