Velita 2kx2k ccd camera

Manufactured by Olympus

The VELITA (2Kx2K) CCD camera is a high-resolution digital imaging device designed for scientific and industrial applications. It features a 2048x2048 pixel CCD sensor, providing detailed image capture capabilities. The camera is optimized for accurate and reliable data acquisition, making it suitable for a wide range of laboratory and research-oriented tasks.

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Lab products found in correlation

Superdex 200, by GE Healthcare (1 mentions) T12 electron microscope, by Olympus (1 mentions) Tecnai 12 biotwin transmission electron microscope, by Thermo Fisher Scientific (1 mentions) Gloqube glow discharge system, by Quorum Technologies (1 mentions)

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CCD camera (126 citations)

4 protocols using velita 2kx2k ccd camera

Negative Stain Electron Microscopy of NFL Trimers

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For negative stain electron microscopy, trimers were purified by lentil lectin affinity chromatography, following which trimer fractions were collected by size exclusion chromatography using a Superdex 200 (GE Healthcare) column pre-equilibrated with buffer containing 5 mM HEPES, 150 mM NaCl (pH 7.5).
The SEC purified samples were prepared for negative staining in the following conditions – (i) NFL Wt, (ii) designed trimer NFL D1, (iii) designed trimer NFL D2, (iv) designed trimer NFL D3. All samples were prepared for Transmission Electron Microscopy imaging by conventional negative staining method. Briefly, 3.5 μl of sample (for all the constructs) was applied to glow discharged carbon coated copper grid for 30 s followed by blotted out excess buffer and sample. Negative staining was performed using 2% uranyl acetate for 25 s. All negative stained samples were visualized and imaged at room temperature using Tecnai T12 electron microscope equipped with a LaB6 filament operated at 120 kV. All images were recorded using a side-mounted Olympus VELITA (2Kx2K) CCD camera at 2.54 Å/pixel on the specimen level.

Malladi S.K., Schreiber D., Pramanick I., Sridevi M.A., Goldenzweig A., Dutta S., Fleishman S.J, & Varadarajan R. (2020). One-step sequence and structure-guided optimization of HIV-1 envelope gp140. Current Research in Structural Biology, 2, 45-55.

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Negative Staining TEM Analysis of S Protein

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S protein samples at three different pH conditions (pH 8.0, pH 7.4 and pH 6.5) were analysed for overall homogeneity and distribution using conventional Negative Staining (NS) TEM. Carbon coated Cu grids (EM grid, 300 mesh, Electron Microscopy Sciences) were glow discharged for 30 sec and 3.5 μl of samples (0.1mg/ml) were added and incubated on the grid for 1.5 min. Excess buffer was blotted and negative staining was performed using 1% uranyl acetate (Uranyl Acetate 98%, ACS Reagent, Polysciences, Inc.). The samples were checked, and data acquisition was performed at room temperature using a 120 kV Tecnai T12 electron microscope. Data were collected at 120 kV using side-mounted Olympus VELITA (2Kx2K) CCD camera calibrated pixel size as 2.96 Å/pixel.

Pramanick I., Sengupta N., Mishra S., Pandey S., Girish N., Das A, & Dutta S. (2021). Conformational flexibility and structural variability of SARS-CoV2 S protein. Structure (London, England : 1993), 29(8), 834-845.e5.

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Negative Staining Nanoparticle Imaging

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All the negative stain EM samples were prepared using staining methods described in previous studies (36 (link), 37 (link)). Initially, glow discharge of the copper grids was carried out for 40-60 seconds at 20 mA current using GloQube glow discharge system (Quorum technologies) for uniform settling of the particles on the grid. Following this, 3.5 µl of nanoparticle (0.4 mg/mL) in 20 mM Tris was adsorbed on the grids for 30-60 secs. Excess buffer was removed by blotting with a filter paper and negative stain was performed using 1% uranyl acetate. Negative stain imaging experiment was carried out at room temperature using a FEI Tecnai 12 BioTwin transmission electron microscope equipped with a LaB6 (lanthanum hexaboride crystal) filament at a voltage of 120 Kv. Datasets were collected using a side-mounted Olympus VELITA (2Kx2K) CCD camera at a calibrated magnification of 75000 x and a defocus value of −1.3 µm. The final images were recorded at a pixel size of 2.54 Å on the specimen level. Following this, properly sized particles were manually chosen and extracted using e2boxer.py and e2projectmanager.py EMAN 2.1 software (38 (link)). Finally, a 2-D reference free classification of particles was carried out using e2refine2d.py.

Kar U., Khaleeq S., Garg P., Bhat M., Reddy P., Vignesh V.S., Upadhyaya A., Das M., Chakshusmathi G., Pandey S., Dutta S, & Varadarajan R. (2022). Comparative Immunogenicity of Bacterially Expressed Soluble Trimers and Nanoparticle Displayed Influenza Hemagglutinin Stem Immunogens. Frontiers in Immunology, 13, 890622.

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Negative Staining for Spike-2P Visualization

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For visualization by transmission electron microscope, Spike-2P sample was prepared by conventional negative staining method. Briefly, carbon-coated Cu grids were glow discharged for 30 s, and 3.5 μl of sample (0.1 mg/ml) was incubated on the grid for 1 min. The extra sample and buffer solution was blotted out, and negative staining was performed using 1% Uranyl Acetate solution for 30 s. Freshly prepared grids were air-dried for 30 min. The negatively stained sample was visualized at room temperature using a Tecnai T12 electron microscope equipped with a LaB₆ filament operated at 120 kV using a low electron dose. Images were recorded using a side-mounted Olympus VELITA (2KX2K) CCD camera using defocus ranging from −1.3 to −1.5 and a calibrated pixel size 2.54 Å/pixel at specimen level.

Malladi S.K., Singh R., Pandey S., Gayathri S., Kanjo K., Ahmed S., Khan M.S., Kalita P., Girish N., Upadhyaya A., Reddy P., Pramanick I., Bhasin M., Mani S., Bhattacharyya S., Joseph J., Thankamani K., Raj V.S., Dutta S., Singh R., Nadig G, & Varadarajan R. (2020). Design of a highly thermotolerant, immunogenic SARS-CoV-2 spike fragment. The Journal of Biological Chemistry, 296, 100025.

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