Frontal Cryosectioning: An Improved Protocol for Sectioning Large Areas of Fibrous Scaffolds

Figure 2

Flattening and freezing a sample for cryosectioning. The flattening process is critically important—ensuring the sample is flush with the bottom of the flat bottomed cryomold allows for accurate alignment of the first, crucial section. Steps (A)–(D) are completed outside of the cryosectioning chamber (room temperature) while steps (E)–(H) are completed inside the cryosectioning chamber, on the cryobar or on an equivalent precooled metal mass. Steps (E) and (F) must be completed quickly after placing the sample in the cryosectioning chamber to prevent the premature freezing of the sample in an unflattened state (e.g., if a sample has residual tension, such as when electrospinning onto a small diameter mandrel, it will tend to curl).