Imaging of Mucin Networks with Atomic Force Microscopy.

Abstract

Mucin networks serve as the structural scaffold of mucus and play a significant role in determining its biophysical properties. Thus, characterizing the organization, macromolecular structure, and interactions within these networks is a key step in understanding the parameters that govern mucus functionality in both health and disease. Atomic force microscopy (AFM) is uniquely suited to study mucin networks; AFM can clearly resolve nanometer-sized features, does not require fixation or metallization, and can be performed in air or aqueous solutions. In this chapter we describe protocols to image mucin networks using AFM. First, we describe two protocols to enrich and isolate mucin samples in preparation for AFM imaging. Next, we detail a protocol to deposit the samples onto a mica substrate. Finally, we give general tips to optimize and troubleshoot AFM imaging of mucin networks.