Cp3 plunge freezing device

Manufactured by Ametek

Sourced in United States

The CP3 plunge freezing device is a compact laboratory instrument designed for the rapid freezing of biological samples. It utilizes a liquid cryogen, such as liquid nitrogen, to achieve ultra-low temperatures for the preservation of sample integrity. The device's core function is to provide a controlled and consistent freezing process for effective sample preparation.

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Lab products found in correlation

Tecnai f20 microscope, by Thermo Fisher Scientific (1 mentions) So 163, by Kodak (1 mentions) Super coolscan 9000 ed scanner, by Nikon (1 mentions)

2 protocols using cp3 plunge freezing device

Cryo-EM Sample Preparation Protocol

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Samples were prepared on a Gatan CP3 plunge freezing device (Gatan, Warrendale, PA, USA). Briefly, 3 μL of sample were applied to glow-discharged Quantifoil grids in an atmosphere of > 80% humidity (Quantifoil, Großlöbichau, Germany). Grids were blotted for 1–2 s then plunged into liquid ethane at − 174 °C. Grids were stored in liquid nitrogen for transport to a Tecnai F20 microscope (Thermo Fisher Scientific, Waltham, MA, USA), and imaged immediately. Images were collected on a Tietz F415 camera (TVIPS, Gauting, Germany) under low dose conditions at 19000X (8.8 A/pixel) using SerialEM software (University of Colorado, Boulder, CO, USA).

Colangelo N.W, & Azzam E.I. (2020). Extracellular vesicles originating from glioblastoma cells increase metalloproteinase release by astrocytes: the role of CD147 (EMMPRIN) and ionizing radiation. Cell Communication and Signaling : CCS, 18, 21.

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Cryo-EM imaging of PCV2 virus

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A 3 μl aliquot of the PCV2 sample at 3 mg/ml concentration was applied to 400-mesh holey carbon grids (1.2/1.3 C-flat, Protochips). After blotting excessive sample solution, the grid was plunge frozen in liquid ethane cooled by liquid nitrogen using the Cp3 plunge-freezing device (Gatan). Cryo-EM images were taken using a FEI Titan Krios electron cryo-microscope with field emission gun (FEG) operated at 300 kV and parallel illumination. The images were recorded on Kodak SO163 negative films at low dose (25–27 e/Å²) and at a nominal magnification of 59k with intended defocus range of 1.5–2.5 μm. The films without obvious ice contamination (141 out of 168) were selected for digitization using a Nikon Super CoolScan 9000ED scanner with step size of 6.35 μm/pixel. The final calibrated sampling of the scanned images is 1.08 Å/pixel.

Liu Z., Guo F., Wang F., Li T.C, & Jiang W. (2016). 2.9 Å Resolution Cryo-EM 3-D Reconstruction of Close-packed Virus Particles. Structure (London, England : 1993), 24(2), 319-328.

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