Figure 2

Slice-and-image operation in cryo-imaging. The diagram illustrates how the cryo-imaging acquires block-face images. a tissue sample (green box), containing a fluorescent cell (yellow circle), is fixed on a cutting stage (blue structures) that can move back and forth between the imaging position and the cutting position. After the first block-face image is taken by the camera above (A), the tissue sample is elevated by a fixed length of X, and then moved toward a sharp knife to cut off the top thin layer (with the thickness of X) (B). After the cut, the sample is moved back to the imaging position so that the second block-face image can be taken (C). The whole process of slice-and-image is repeated (D–E) until the whole sample is gone. Please note that this diagram is not to scale.