Analyzing the distribution of cell membrane receptors in real-time based on multiple molecule force spectroscopy

Most cellular activities are predicated on ligand-receptor interactions at the cell membrane, the distribution and expression level of receptors on cell membrane exhibit significant variability. In this study, we introduce a new method to analyze the distribution of cell membrane receptors in real-time. Herein, the multiple molecule force spectroscopy (MMFS) was employed by using ligands functionalized microsphere on AFM (atomic force microscopy) tip cantilever. Based on the MMFS, the cell membrane distribution of folate receptor (FR) and epidermal growth factor receptor (EGFR) on HeLa cell, A549 cell, and Vero cell is evaluated. Meanwhile, the distribution difference of the receptors on perinuclear and nuclear of cell membrane was identified. Furthermore, this method is used to monitor the EGFR distribution on cell membrane with the regulation of hyaluronic acid (HA) with different disaccharide units in real time. The enhancement effect of HA with different disaccharide number on the expression level of EGFR was quantified. This method holds promise for facilitating regulation the cell membrane proteins in real-time during various complex biological activities.