Super-resolution imaging of folate receptor alpha on cell membranes using peptide-based probes

Folate receptor alpha (FRα) is a vital membrane protein which have great association with cancers and involved in various biological processes including folate transport and cell signaling. However, the distribution and organization pattern of FRα on cell membranes remains unclear. Previous studies relied on antibodies to recognize the proteins. However, multivalent crosslinking and large size of antibodies confuse the direct observation to some extent. Fortunately, the emergence of peptide, which are small-sized and monovalent, has supplied us an unprecedented choice. Here, we applied fluorophore-conjugated peptide probe to recognize the FRα and study the distribution pattern of FRα on cell membrane using dSTORM super-resolution imaging technique. FRα were found to organized as clusters on cell surface with different sizes. And they have a higher expression level and formed larger clusters on various cancer cells than normal cells, which hinted that its specific distribution could be utilized for cancer diagnosis. Furthermore, we revealed that the lipid raft and cortical actin as restrictive factors for the FRα clustering, suggesting a potential assembly mechanism insight into FRα clustering on cell membrane. Collectively, our work clarified the morphology distribution and clustered organization of FRα with peptide probes at the nanometer scale, which paves the way for further revealing the relationship between the spatial organization and functions of membranal proteins.