Sample preparation for TEM imaging was based on the protocol described by Castro et al.42 (link). Briefly, 12 μL of the sample solutions (~10 nM concentration) were placed on glow-discharged carbon-coated 400 mesh copper TEM grids. After 2 min, the samples were wicked off of the grid with filter paper and immediately replaced with 12 μL of freshly prepared uranyl formate negative staining solution. After 30 sec, the stain was removed and the grids were allowed to air dry. Images were acquired at 25,000x using a JEOL 1230 TEM (Peabody, MA) equipped with a Gatan Inc. 2k × 2k Ultrascan camera (Pleasanton, CA).
Ultrascan 2k 2k camera
Manufactured by Ametek
Sourced in United States
The UltraScan 2k × 2k camera is a high-resolution imaging device designed for laboratory applications. It features a 2048 × 2048 pixel sensor, providing a detailed and accurate capture of samples or specimens under investigation.
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Lab products found in correlation
7 protocols using ultrascan 2k 2k camera
1
Negative Staining of Biomolecules for TEM
2
Transmission Electron Microscopy of Nanoparticles
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TEM was performed as previously described25 (link). Briefly, samples for TEM imaging were prepared in a concentration range of 0.5 nM to 5 nM. 12 μL of the sample was placed on glow-discharged carbon-coated 400 mesh copper TEM grids. After two minutes of incubation, the sample solution was removed using filter paper and replaced with 12 μL of freshly prepared uranyl formate negative staining solution. The stain was removed after 30 s, and the grids were air-dried. TEM images were acquired at 25,000 × magnification using a JEOL 1230 TEM (Peabody, Ma, USA) equipped with a Gatan Inc. 2 k × 2 k Ultrascan camera (Pleasanton, CA, USA)25 (link).
3
Negative Stain TEM Imaging of EVs
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EVs were examined by transmission electron microscopy negative stain at the University of Florida Interdisciplinary Center for Biotechnology Research (ICBR) Electron Microscopy Core, RRID: SCR_019146. Poly-l -lysine–treated, 400-mesh carbon-coated Formvar copper grid was floated onto 3 μl of aliquoted EV suspension for 5 min, incubated on 2% paraformaldehyde in PBS (pH 7.20) for 15 min, followed by a PBS wash and water wash for 5 min, each. Excess solution was drawn off with filter paper, and the grid was floated onto 1% aqueous uranyl acetate for 30 s. The stain was removed with filter paper, air-dried, and examined with a FEI Tecnai G2 Spirit Twin TEM (FEI Corp., Hillsboro, OR) operated at 120 kV. Digital images were acquired with a Gatan UltraScan 2k × 2k camera and Digital Micrograph software (Gatan Inc., Pleasanton, CA).
4
Negative Staining Electron Microscopy of Tau Fibrils
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Negative staining electron microscopy was performed on tau fibrils. 2 μL of tau fibrils (1 mg/mL) were applied to 300 mesh carbon coated copper grids (Electron Microscopy Sciences, Hatfield, PA) and were allowed to settle for 10 min. Grids were washed with filtered PBS. 1% uranyl acetate was filtered through a 2-micron filter and applied to each grid that were then dried. Samples were examined with a FEI Tecnai G2 Spirit Twin TEM (FEI Corp., Hillsboro, OR) operated at 120 kV and digital images were acquired with a Gatan UltraScan 2 k × 2 k camera and Digital Micrograph software (Gatan Inc., Pleasanton, CA).
5
Ultrastructural Analysis of Rat Islets
6
Transmission Electron Microscopy of Porphyromonas gingivalis Biofilm
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Forty microliters of 1x PBS was placed on a single P. gingivalis colony biofilm and suspended. Poly-l -lysine treated, glow discharged, carbon-coated Formvar 400 mesh copper grid was floated onto the suspended colony for 5 min, followed by a quick water wash. Without letting the grid dry, the grid was floated onto a 10 µl droplet of 4% paraformaldehyde for 5 min. Excess solution was dabbed off with filter paper and grid was floated on 0.5% aqueous uranyl acetate for 30 s. Stain was removed with filter paper, air dried and examined with FEI Tecnai G2 Spirit Twin TEM (FEI Corp.) and digital images were acquired with Gatan UltraScan 2k × 2k camera and Digital Micrograph software (Gatan Inc.).
7
Ultrastructural Analysis of Human Liver
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Explanted human liver tissue was processed and imaged at the University of Florida ICBR Electron Microscopy Core Facility. For transmission electron microscopy (TEM), pieces of human liver explant were fixed in 4% paraformaldehyde and 2.5% glutaraldehyde. Dehydrated samples were infiltrated in anhydrous acetone-Spurr’s epoxy resin with Z6040 embedding primer (Electron Microscopy Sciences) and semi-thick sections were collected. Prefixed tissues were then trimmed and immunogold labeling was performed using polyclonal rabbit anti-AAT antibody (Dako) at 1:10 dilution. Ultrathin sections were examined with a FEI Tecnai G2 Spirit Twin TEM (FEI Corp.) operated at 120 kV and digital images were acquired with a Gatan UltraScan 2k×2k camera and Digital Micrograph software (Gatan Inc.).
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