Abstract: Lipids play essential biological functions that include acting as components of biological membranes, energy storage, signaling, nutrients, transporters, enzyme activators, among others. Compared with the multiple research methods to assess DNA, RNA, and protein content, location, and function in cells, there are relatively fewer methods to study lipids. Therefore, lipid-oriented mechanistic studies remain rare and challenging. Lipidomics which allows large-scale analysis of cellular lipids, is a critical strategy for achieving this. One revolutionary advance in lipidomics pioneered by our group is the development of multidimensional mass spectrometry-based shotgun lipidomics, which has become a foundational analytical technology platform among current lipidomics practices due to its high efficiency, sensitivity, and reproducibility, as well as its broad coverage and minimal batch effects.