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

Figure 4

Consecutive frontal images of electrospun scaffolds seeded with human dermal fibroblasts stained with DAPI (yellow). Note how data management can be monitored by examining sections prior to image acquisition. Once some predetermined threshold of cells has been detected in the scaffold frontal sections the data sets are prepared for -stack assembly. In this example, the majority of cells were present in the first 70 μm of the sample and subsequent frontal sections were excluded from the analysis. The large scale pores present in this scaffold were used to align the samples in the vertical direction prior to -stack assembly.