EM was performed at the CMM Electron Microscopy Facility at University of California San Diego. Four-month organoids were immersed in modified Karnovsky’s fixative (2.5 % glutaraldehyde and 2 % paraformaldehyde in 0.15 M sodium cacodylate buffer, pH 7.4) for at least 4 hours, post fixed in 1% osmium tetroxide in 0.15 M cacodylate buffer for 1 hour and stained in 2 % uranyl acetate for 1 hour. Samples were dehydrated in ethanol, embedded in Durcupan epoxy resin (Sigma-Aldrich), sectioned at 50 to 60 nm on a Leica Ultracut UCT (Leica, Bannockburn, IL), and transfer onto Formvar and carbon-coated copper grids. Sections were stained with 2 % uranyl acetate for 5 minutes and Sato's lead stain for 1 minute. Grids were analyzed using a JEOL 1200EX II (JEOL, Peabody, MA) transmission electron microscope equipped with a Gatan digital camera (Gatan, Pleasanton, CA).
Gatan digital camera
Manufactured by Ametek
Sourced in United States
The Gatan digital camera is a high-performance imaging device designed for use in electron microscopy. It captures digital images of samples under observation in the microscope.
Automatically generated - may contain errors
Lab products found in correlation
Durcupan epoxy resin, by Merck Group (5 mentions)
1200 ex 2, by JEOL (5 mentions)
Uct ultramicrotome, by Leica (5 mentions)
Ultracut uct, by Leica (1 mentions)
Uranyl acetate, by Ladd Research Industries (1 mentions)
Orius 600 digital camera, by Ametek (1 mentions)
1400 transmission electron microscope, by JEOL (1 mentions)
Ultracut uct ultramicrotome, by Leica (1 mentions)
Jem 1400 electron microscope, by JEOL (1 mentions)
Tecnai g2 spirit biotwin tem, by Thermo Fisher Scientific (1 mentions)
1200 ex 2 tem, by JEOL (1 mentions)
Eagle 4k hs digital camera, by Thermo Fisher Scientific (1 mentions)
Jem 1400 transmission electron microscope, by JEOL (1 mentions)
12 protocols using gatan digital camera
1
Ultrastructural Analysis of Organoid Samples
2
TEM Analysis of Extracellular Vesicles and Liver Samples
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For transmission electron microscopy, extracellular vesicles were adhered to 100 mesh Formvar and carbon coated grids for 5 minutes at room temperature. Grids were washed once with water, stained with 1% uranyl acetate (Ladd Research Industries, Williston VT) for 1 minute, dried and viewed using a JEOL 1200 EXII transmission electron microscope. Images were captured using a Gatan Orius 600 digital camera (Gatan, Pleasanton, CA). Liver samples were collected from the CDAA-fed mice after a short liver perfusion with 10 mL of 4% paraformaldehyde in 0.15 M sodium cacodylate buffer, pH 7.4 by using a 21 G needle. Samples were immersed in modified Karnovsky’s fixative (2.5% glutaraldehyde and 2% paraformaldehyde in 0.15 M sodium cacodylate buffer, pH 7.4) for at least 4 hours, postfixed in 1% osmium tetroxide in 0.15 M cacodylate buffer for 1 hour and stained en bloc in 3% uranyl acetate for 1 hour. Samples were dehydrated in ethanol, embedded in Durcupan epoxy resin (Sigma-Aldrich), sectioned at 50 to 60 nm on a Leica UCT ultramicrotome, and picked up on Formvar and carbon-coated copper grids. Sections were stained with 3% uranyl acetate for 5 minutes and Sato’s lead stain for 1 minute. Grids were viewed using a JEOL 1200EX II (JEOL, Peabody, MA) transmission electron microscope and photographed using a Gatan digital camera (Gatan, Pleasanton, CA).
3
Ultrastructural Analysis of Vc/C2 in CCI-ION Rats
4
Ultrastructural Analysis of Coral Tissue
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Corals were fixed overnight in modified Karnovsky’s fixative (2.5% glutaraldehyde and 2% paraformaldehyde in 0.15 M sodium cacodylate buffer, pH 7.4). Coral fragments were then decalcified and post-fixed in 1% osmium tetroxide in 0.15 M cacodylate buffer for 1 hour and stained en bloc in 2% uranyl acetate for 1 hour. Samples were dehydrated in ethanol, embedded in Durcupan epoxy resin (Sigma-Aldrich, St. Lewis, MO, USA), sectioned at 50 to 60 nm on a Leica UCT ultramicrotome (Leica, Bannockburn, IL, USA), and picked up on Formvar and carbon-coated copper grids. Sections were stained with 2% uranyl acetate for 5 minutes and Sato’s lead stain for 1 minute. Grids were viewed using a JEOL 1200EX II (JEOL, Peabody, MA, USA) transmission electron microscope and photographed using a Gatan digital camera (Gatan, Pleasanton, CA, USA).
5
Cell Morphology Analysis using Microscopy
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Cell morphology was observed using Olympus microscope BX51 (Tokyo, Japan) fitted with a CapturePro 2.6-JENOPTIK Laser, Optik, System (GmbH, Germany) camera and an FEI Tecnai transmission electron microscope at an accelerating voltage of 80 kV. Electron micrographs were taken using a Gatan digital camera (Gatan, Oxon, UK).
6
Ultrastructural Analysis of Muscle Fibers
7
Transmission Electron Microscopy of Micellar Structures
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The morphology of self-assembled structures under study was investigated by TEM using a Morgagni TEM (Field Emission Inc., Hillsboro, OR, USA) with a Gatan digital camera (Gatan, Pleasanton, CA, USA). Furthermore, 20 μL of the micellar solution with a polymer concentration of 1 mg·mL−1 was placed on a copper-coated grid. The grid was held horizontally for 1 min to allow the colloidal particles to settle down. The excess fluid was removed by filter paper. The copper-coated grids holding the aqueous samples were then negatively stained by 2% phosphotungstic acid. After 2 min, the excess fluid was removed by filter paper and the grid was loaded into TEM for image analysis.
8
Transmission Electron Microscopy of Self-Assembled Structures
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The morphology of self-assembled structures under study was investigated by TEM using a Morgagni TEM (Field Emission Inc., Hillsboro, OR, USA) with Gatan digital camera (Gatan, Pleasanton, CA, USA). In brief, 20 μL of micellar solution with a polymer concentration of 0.25 mg/mL or Cremophor EL at a concentration of 0.2 mg/mL was placed on a copper-coated grid. The grid was held horizontally for 1-2 min to allow the colloidal particles to settle down. The excess fluid was removed by filter paper. The copper-coated grids holding the aqueous samples were then negatively stained by 2% phosphotungstic acid. After 2 min, the excess fluid was removed by filter paper and the grid was loaded into the TEM for image analysis.
9
Electron Microscopy Imaging of Thioflavin T Aggregates
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To observe the morphology of aggregate species used in the thioflavin T binding assay, electron micrographs were prepared for each of the samples described above. A 10 μL drop of the sample was placed on a sheet of parafilm and a copper mesh electron microscopy grid (Electron Microscopy Sciences, Hatfield, PA, USA) was incubated with the sample for 2 min. Following incubation and removal of excess liquid, 10 μL of 1% uranyl acetate solution was placed on the grid and incubated for 30 s. Upon removal of excess liquid, the grid was washed with 10 μL of 1% uranyl acetate and dH2O, and was dried for 1 h. Grids were imaged using a JEOL JEM-1400 electron microscope (JEOL, Peabody, MA, USA) at 100 kV and photographed using a Gatan digital camera (Gatan Inc., Pleasanton, CA, USA).
10
Transmission Electron Microscopy of NSCs
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Transmission electron microscopy was performed human and mouse NSCs. Samples were immersed in modified Karnovsky's fixative (2.5% glutaraldehyde and 2% paraformaldehyde in 0.15 M sodium cacodylate buffer, pH 7.4) for at least 4 h, post‐fixed in 1% osmium tetroxide in 0.15 M cacodylate buffer for 1 h, and stained en bloc in 2% uranyl acetate for 1 h. Samples were dehydrated in ethanol, embedded in Durcupan epoxy resin (Sigma‐Aldrich), sectioned at 50–60 nm using a Leica UCT ultramicrotome, and picked up on Formvar and carbon‐coated copper grids. Sections were stained with 2% uranyl acetate for 5 min and with Sato's lead stain for 1 min. Grids were viewed with (i) a JEOL 1200EX II TEM (JEOL, Peabody, MA) and photographed using a Gatan digital camera (Gatan, Pleasanton, CA) or (ii) a Tecnai G2 Spirit BioTWIN TEM equipped with an Eagle 4k HS digital camera (FEI, Hilsboro, OR).
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