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. 2016 Feb 10;7:60. doi: 10.3389/fpls.2016.00060

Figure 2.

Figure 2

Adhesive film assisted CMC embedding method. (i) Freeze embedding can be achieved by either (1) freezing before embedding or (2) freezing after embedding. In freezing before embedding method: (a,b) Freezing of the sample in the coolant; (c) freeze-embedding of the frozen sample with 4–5% CMC gel; (d) completely freezing the CMC gel in coolant (i.e., hexane); (e) removing the CMC block from container;. In freezing after embedding method: (a) placing the sample into 4–5% CMC gel; (b) completely freezing the CMC gel in coolant (i.e., hexane); (c) removing the CMC block from container; (ii) After freeze embedding, the CMC block is subjected to freeze sectioning: (f) fixing the embedded sample to the sample holder with CMC gel and then attaching it to the cryomicrotome; (g) adhering the film to the exposed cutting surface; (h) cutting the sample; (i) placing sample section on the glass slide. Figure is adapted with permission from Kawamoto (2003).