Cell adhesion is of fundamental importance in the cell communication, signaling and motility, and its dysfunction occurs prevalently during cancer progression. The knowledge of the molecular and cellular processes involved in abnormalities in cancer cells adhesion has dramatically increased and it has been focused mainly on cellular adhesion molecules (CAMs) and tumor microenvironment. During the last decade, several methods for measuring and studying cell adhesion properties were also developed, including optical tweezers technology. We have developed a non-invasive method employing optical tweezers technology in order to differentiate normal B-cells and non-Hodgkin’s lymphoma cells derived from clinical samples. Our approach bases on nascent adhesion formation between B-cell and mesenchymal stromal cell. In this method, a single B-cell is trapped and optically seeded on mesenchymal stromal cell and maintained direct contact until stable connection between cells is formed in time-scale. After we characterized the adhesive properties of the panel of clinical samples, we performed the deep proteomics investigations on cellular adhesion proteins using liquid chromatography- tandem mass spectrometry (LC-MS/MS).