Extended Data Fig. 3: Workflow of 3D correlative cryo-FIB and cryo-ET. | Nature Microbiology

Extended Data Fig. 3: Workflow of 3D correlative cryo-FIB and cryo-ET.

From: HIV-1 nuclear import is selective and depends on both capsid elasticity and nuclear pore adaptability

Extended Data Fig. 3

a, Illustration of 3D correlative cryo-FIB milling workflow. Yellow circles with numbers are ice particles used as fiducial markers for the correlation. Three white arrowheads point to targeted HIV-1 cores. (i) Fluorescence image of the targeted nucleus after rough milling, maximum intensity projection (MIP) from 31 images, step size = 500 nm, the region of interest (ROI) is enlarged in the inset; (ii) Cryo-FIB image of the targeted nucleus after rough milling; (iii) Cryo-FIB image of the polished lamella; (iv) Fluorescence image of the polished lamella, the ROI is enlarged in the inset, MIP from 151 images, step size = 100 nm. b-d, Demonstration of the correlative imaging from cryo-FLM to cryo-FIB and to cryo-EM, as shown with MIP fluorescence images before (b) and after (c) milling and overlapped with the TEM overview of the same lamella (d). Scale bars = 5 μm.

Back to article page