Novel techniques for combined MALDI-FRAP measurements and mapping

Novel approaches for the measurements of lateral diffusion / molecular mobility and bindings using MALDI + FRAP and MALDI + FLIP hybridization are proposed (“MALDI-FLIP-on-a-chip” and “MALDI-FRAP-on-a-flap”). Novel MALDI MS + FLIP approaches for verifying continuity of membranous structures and measurements of nucleus-cytoplasm exchange rates are proposed. DEFINITIONS: FRAP (Fluorescence Recovery After Photobleaching) is a method for diffusion kinetics measurements in living cells using fluorescence microscopy which allows to estimate quantitatively 2D lateral diffusion in molecularly thin film containing fluorescent-labeled probes, or for single cell examination (i.e. the study of lateral mobility of cellular molecules). FLIP (Fluorescence Loss in Photobleaching) is a microscopic technique predominantly performed using laser scanning microscopy (e.g. for tagged protein local photobleaching by short, intensive laser excitation on CLSM platform) used for the studies on molecular mobility inside the cells and membranes. MALDI (Matrix-Assisted Laser Desorption/Ionization) is a soft ionization technique used in mass spectrometry, allowing for the analysis of biomolecules.