Quantification of Phosphatidylserine Molecules on the Surface of Individual Cells Using Single-Molecule Force Spectroscopy

Abstract

Identification of the phosphatidylserine (PS) discrepancies occurring on the cellular membrane during apoptotic processes is of the utmost importance. However, monitoring the quantity of PS molecules in real-time at a single-cell level currently remains a challenging task. Here, we demonstrate this objective by leveraging the specific binding and reversible interaction exhibited by the zinc(II) dipyridinamine complex (ZnDPA) with PS. Lipoic acid-functionalized ZnDPA (LP-ZnDPA) was subsequently immobilized onto the surface of an atomic force microscopy cantilever to form a force probe, ALP−ZnDPA, enabling a PS-specific dynamic imaging and detection mode. By utilizing this technique, we can not only create a heat map of the expression level of PS with submicron resolution but also quantify the number of molecules present on a single cell's surface with a detection limit of 1.86 × 10 4 molecules. The feasibility of the proposed method is demonstrated through the analysis of PS expression levels in different cancer cell lines and at various stages of paclitaxel-induced apoptosis. This study represents the first application of a force probe to quantify PS molecules on the surface of individual cells, providing insight into dynamic changes in PS content during apoptosis at the molecular level and introducing a novel dimension to current detection methodologies.

DHM/SERS reveals cellular morphology and molecular changes during iPSCs-derived activation of astrocytes

“…SERS detected a decrease in phospholipid-related spectral peaks after astrocyte activation, including phosphatidylserine at 524 cm − 1 , phosphatidylinositol at 776 cm − 1 , and phospholipids at 1745cm − 1 . Phosphatidylserine, an aminophospholipid located in the inner leaflet of the membrane, is closely related to inflammatory responses, oxidative stress, and apoptosis [ 44 ]. Previous reports have indicated that changes in phospholipids suggest alterations in the composition of glial cell membranes during neuroinflammation [ 45 ].…”