Figure 1.

Cryo-FM Allows Selection of Vitreous Section Areas that Are Suitable for Cryo-ET and Contain Fluorescent Signals of Interest

(A) Grid overview in the green channel shows the distribution of vitreous sections appended to each other as ribbons.

(B and C) Two planes from the same focal stack, separated by about 3.6 μm in z (direction of the light path), merge of red (shown in magenta) and green channels. Area of acquisition corresponds to the yellow dashed box in (A). The white box in (B) indicates a section area where the holey carbon film and the GFP signals in the section are in focus in the same plane. The magenta box indicates a section area where the carbon film is in focus in the plane shown in (B), whereas the GFP signals are in focus in a different plane shown in (C).

(D) Intermediate-magnification EM image of the area indicated by the white dashed square in (B), magnified in (E).

(E) The white circle marks a Pil1-GFP signal suitable for cryo-ET acquisition. The orange circles in (D and E) mark examples of fiducial markers.

(F) Intermediate-magnification EM image of the area indicated by the magenta dashed square in (B and C), magnified in (G).

(G) The white circle indicates a Pil1-GFP spot not suitable for cryo-ET acquisition.

Scale bars represent 100 μm in (A), 10 μm in (B and C), 500 nm in (D and F).