The negatively stained specimen was examined on a Zeiss Libra 120 Plus TEM (Carl Zeiss NTS, Oberkochen, Germany) operating at 120 kV with 20 eV in-column energy filtering at room temperature. Micrographs were acquired by a Gatan UltraScan 4Kx4K CCD at 80,000x magnification (each pixel corresponding to 1.48 Å) under near Scherzer focus (0.1 μm) and defocus of 0.4 μm. Micrographs were processed with EMAN, SPIDER, and FREALIGN software packages as described elsewhere [17 – 20 (link)]. The contrast transfer function parameters of each micrograph were determined and corrected. A total of 2,986 rHDL were used for reference-free class averaging with the EMAN software package.
Ultrascan 4kx4k ccd
Manufactured by Ametek
The UltraScan 4Kx4K CCD is a high-resolution charge-coupled device (CCD) imaging sensor. It features a 4,096 x 4,096 pixel array, enabling the capture of large, detailed images. The CCD is designed for use in advanced scientific and industrial applications that require high-quality image data.
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Lab products found in correlation
2 protocols using ultrascan 4kx4k ccd
1
Structural Analysis of Reconstituted HDL
2
Tilt Series Cryo-TEM Imaging Protocol
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The TEM holder was tilted at angles ranging from −45° to +45° in 1.5° increments, and imaging was controlled using Gatan tomography software (Zeiss Libra 120 Plus TEM). The TEM was operated at 120 kV high tension with a 20-eV energy filter. The tilt series was acquired under low defocus conditions (<1 μm) using a Gatan UltraScan 4 K X 4 K CCD under a magnification of 80 kx (each pixel of the micrographs corresponds to 1.48 Å in specimens). The total electron doses were 2,800–6,600 e−Å−2. The micrographs were initially aligned using the IMOD103 (link) software package. The CTF was then corrected using TomoCTF104 (link). The tilt series of the particles in square windows of 256 × 256 pixels (~38 nm) were semi-automatically tracked, windowed using IPET software39 (link), and finally binned by 2 to reduce computation time in the subsequent reconstruction.
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