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Ultracam

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The UltraCam is a high-performance digital camera system designed for aerial photography and remote sensing applications. It captures high-resolution, multispectral imagery with exceptional image quality and radiometric performance.

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Lab products found in correlation

Vitrobot, by Thermo Fisher Scientific (2 mentions) Polara electron microscope, by Thermo Fisher Scientific (2 mentions) Tecnai f30, by Thermo Fisher Scientific (2 mentions) Feg tem, by Thermo Fisher Scientific (1 mentions)

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4k × 4k UltraCam (2 citations)

5 protocols using ultracam

1

Purification and Imaging of Yeast Spindle Pole Bodies

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Spindle pole bodies from Saccharomyces cerevisiae were purified following previous published procedures 34 ,35 . The purified SPB sample, initially in high concentration of sucrose, was first dialyzed at 4°C overnight in a buffer containing 10 mM Bis-Tris/Cl (pH=6.5), 0.1 mM MgCl2, 20% (v/v) DMSO. Next day, after mixing with 10 nm colloid gold, the sample was applied onto either a home-made holey carbon grid or a Quantifoil grid (PSI, Inc.) in a humidity chamber, then blotted and plunged into liquid ethane using a home-made plunger or a Vitrobot (FEI, Inc.). Frozen grids were stored in liquid nitrogen before use. Tomography data were collected on a Polara electron microscope (FEI, Inc.) running at 300kV. A post-column energy filter (GIF, Gatan, Inc.) was used and the slit width was set at 25 eV. Automatic data collection was carried out by UCSF Tomography software 36 . Single-axis tilt series were collected at a nominal magnification of 41,000. Images of dimension 2032x2032 were recorded on a CCD camera (UltraCam, Gatan, Inc.). The final pixel size on the images was 5.32 Å. The specimen was tilted from −60° to +60° in 1.5° step. The microscope defocus values were set in the range of 10 to 15 μm. The accumulated dose for each tilt series was ~60 e−/Å2.
Kollman J.M., Greenberg C.H., Li S., Moritz M., Zelter A., Fong K.K., Fernandez J.J., Sali A., Kilmartin J., Davis T.N, & Agard D.A. (2015). Ring closure activates yeast γTuRC for species-specific microtubule nucleation. Nature structural & molecular biology, 22(2), 132-137.
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2

3D Reconstruction of Cryo-TEM Tilt Series

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Single-axis tilt series from −60° to 60° with images were collected in 1° increments and an underfocus of 12 μm using a 300 keV FEI Polara FEG TEM controlled by Leginon software (Suloway et al. 2009 (link)) and the cumulative dose was not allowed to exceed 200 e Å2. The images were recorded on a 4096 × 4096 pixel Ultracam (Gatan, Pleasanton, CA) at a magnification of 22,500 (0.98 nm/pixel). The IMOD software package (Kremer et al. 1996 (link)) was used to calculate 3D reconstructions.
Müller A., Beeby M., McDowall A.W., Chow J., Jensen G.J, & Clemons W.M. (2014). Ultrastructure and complex polar architecture of the human pathogen Campylobacter jejuni. MicrobiologyOpen, 3(5), 702-710.
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3

Purification and Imaging of Yeast Spindle Pole Bodies

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Spindle pole bodies from Saccharomyces cerevisiae were purified following previous published procedures 34 ,35 . The purified SPB sample, initially in high concentration of sucrose, was first dialyzed at 4°C overnight in a buffer containing 10 mM Bis-Tris/Cl (pH=6.5), 0.1 mM MgCl2, 20% (v/v) DMSO. Next day, after mixing with 10 nm colloid gold, the sample was applied onto either a home-made holey carbon grid or a Quantifoil grid (PSI, Inc.) in a humidity chamber, then blotted and plunged into liquid ethane using a home-made plunger or a Vitrobot (FEI, Inc.). Frozen grids were stored in liquid nitrogen before use. Tomography data were collected on a Polara electron microscope (FEI, Inc.) running at 300kV. A post-column energy filter (GIF, Gatan, Inc.) was used and the slit width was set at 25 eV. Automatic data collection was carried out by UCSF Tomography software 36 . Single-axis tilt series were collected at a nominal magnification of 41,000. Images of dimension 2032x2032 were recorded on a CCD camera (UltraCam, Gatan, Inc.). The final pixel size on the images was 5.32 Å. The specimen was tilted from −60° to +60° in 1.5° step. The microscope defocus values were set in the range of 10 to 15 μm. The accumulated dose for each tilt series was ~60 e−/Å2.
Kollman J.M., Greenberg C.H., Li S., Moritz M., Zelter A., Fong K.K., Fernandez J.J., Sali A., Kilmartin J., Davis T.N, & Agard D.A. (2015). Ring closure activates yeast γTuRC for species-specific microtubule nucleation. Nature structural & molecular biology, 22(2), 132-137.
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4

Cryo-Electron Microscopy Protocols for Structural Analysis

Cited 2 times
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Cryo-electron microscopy was performed as in (35 (link),36 (link)). In brief, images (electron dose of ~25 e−/Å2;−4 to−6 µm defocus) were acquired at 27,500×on a Tecnai F30 electron microscope (FEI Company Ltd.) operated at 300 kV. The detector was a GATAN UltraCam, lens-coupled, 4 K CCD camera (binned by 2) attached to a Tridiem Gatan Image Filter (GIF: operated at zero-loss mode with an energy window of 20 eV; Gatan, Inc., Pleasanton, CA, USA). For cryo-electron tomography, tilt series were acquired at−4 or−6 µm defocus, every 1.5 degree (±60 degrees) using serialEM software (37 (link)). The total electron dose was kept at 80–120 e−/Å2.
Höög J.L, & Lötvall J. (2015). Diversity of extracellular vesicles in human ejaculates revealed by cryo-electron microscopy. Journal of Extracellular Vesicles, 4, 10.3402/jev.v4.28680.
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5

Cryo-electron Tomography of TAILS

Cited 1 time
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Microscopy was performed as described previously46 (link),54 . In brief, images (electron dose of ~25 e−/Å2; −4 to −6 µm defocus) were acquired at 27500 x on a Tecnai F30 electron microscope (FEI Company Ltd) operated at 300 kV. The detector was a GATAN UltraCam, lens-coupled, 4 K CCD camera (binned by 2) attached to a Tridiem Gatan Image Filter (GIF: operated at zero-loss mode with an energy window of 20 eV; Gatan Inc., Pleasanton, CA, USA). For tomography, tilt-series were acquired every 1.5 degree (±60 degrees) using serialEM software55 (link). The total electron dose was kept between 80–120 e−/Å2. Cryo-ETs were calculated using eTomo and CTF correction was applied to all datasets. All tomograms were acquired on cells from a single donor and cryo-electron micrographs were acquired from the other two donors to confirm the presence of TAILS.
Zabeo D., Heumann J.M., Schwartz C.L., Suzuki-Shinjo A., Morgan G., Widlund P.O, & Höög J.L. (2018). A lumenal interrupted helix in human sperm tail microtubules. Scientific Reports, 8, 2727.
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