We have previously developed analytical methods for the characterization of trace phospholipids from selectively sampled single cells and intracellular organelles. [1,2] By combining precise micro-sampling techniques with highly sensitive mass spectrometric analysis, these approaches enable molecular investigations at the single-cell and subcellular levels, providing insights into cellular heterogeneity and organelle-specific lipid composition. In this presentation, we will introduce an integrated workflow consisting of SS2000-based single-cell sampling, confocal image analysis, and high-sensitivity direct-infusion nano-electrospray ionization mass spectrometry (nanoESI-MS). The unique features of each technology will be described, including the ability of the SS2000 system to selectively collect target cells or intracellular structures while preserving spatial information obtained through microscopic observation. Furthermore, we will present representative application examples demonstrating the detection and profiling of phospholipids from individual cells and organelles. These results highlight the potential of this analytical platform for investigating cellular heterogeneity, intracellular lipid dynamics, and molecular mechanisms that are difficult to elucidate using conventional bulk-cell analytical approaches.