4k 4k ceta ccd camera

The purified crosslinked WVZ complex was applied to Superose 6 Increase 10/300 GL column (GE Healthcare) equilibrated with the gel filtration buffer. The complex was diluted to 35 ng/μL with SEC buffer. Five microliters of sample were applied to the glow-discharged 200 mesh carbon-coated copper grids. The samples were stained using 0.75% uranyl formate and air-dried. Data were collected on a Talos L120C transmission electron microscope equipped with a 4K × 4K CETA CCD camera (Thermo Fisher Scientific). Images were recorded at a nominal magnification of 92,000×, corresponding to a pixel size of 1.55 Å. CTF parameters of each micrograph were determined using CTFFIND4.^{39 (link)} Particles were picked and subjected to two rounds of 2D classification, followed by de novo 3D model generation, 3D classification, and auto-refinement in Relion.^{37 (link)}

The impact of CSN1201 deletion on the morphology of C. neoformans was assessed by TEM. Cryptococcal cells of both the wild-type (WT) and csn1201Δ strain were cultured to the logarithmic phase in YPD (30°C) or DMEM (37°C, 5% CO₂). The cells were harvested and fixed as previously described (19 (link)). Ultrathin sections (approximately 70 nm thick) were generated using the EM UC7 ultramicrotome (Leica, Wetzlar, Germany), collected on copper grids, and stained with 2% uranyl acetate and Sato’s triple lead stain. All sections were imaged on a Talos L120C TEM (Thermo Fisher Scientific, Waltham, MA, USA) at 80 kV equipped with a 4k×4k Ceta CCD camera.