Alcian blue staining to visualize intracellular hyaluronic acid-based nanoparticles

Investigating at transmission electron microscopy the intracellular trafficking of hyaluronic acid-based nanoparticles remains a challenge due to their intrinsic weak electron density. Here we describe a simple protocol to stain hyaluronic acid that allows visualization of hyaluronic acid-based nanoparticles inside cells at both light and electron microscopy. By applying the critical-electrolyte-concentration Alcian blue method, these nanoparticles were observed as blue dots at bright-field microscopy or filled with fine electron dense precipitates at transmission electron microscopy.