On-Tissue Derivatization of Lipopolysaccharide for Detection of Lipid A Using MALDI-MSI

Hyojik Yang, Courtney E. Chandler, Shelley N. Jackson, Amina S. Woods, David R. Goodlett, Robert K. Ernst, Alison J. Scott*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review


We developed a method to directly detect and map the Gram-negative bacterial virulence factor lipid A derived from lipopolysaccharide (LPS) by coupling acid hydrolysis with matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI). As the structure of lipid A (endotoxin) determines the innate immune outcome during infection, the ability to map its location within an infected organ or animal is needed to understand localized inflammatory responses that results during host-pathogen interactions. We previously demonstrated detec- tion of free lipid A from infected tissue; however detection of lipid A derived from intact (smooth) LPS from host-pathogen MSI studies, proved elusive. Here, we detected LPS-derived lipid A from the Gram-negative pathogens, Escherichia coli (Ec, m/z 1797) and Pseudomonas aeruginosa (Pa, m/z 1446) using on-tissue acid hydrolysis to cleave the glycosidic linkage between the polysaccharide (core and 0-antigen) and lipid A moieties of LPS. Using accurate mass methods, the ion corresponding to the major Ec and Pa lipid A species (m/z 1797 and 1446, respectively) were unambiguously discriminated from complex tissue substrates. Further, we evaluated potential delocalization and signal loss of other tissue lipids and found no evidence for either, making this LPS-to-Lipid A-MSI (LLA-MSI) method, compatible with simultaneous host-pathogen lipid imaging following acid hydrolysis. This spatially sensitive technique is the first step in mapping host-influenced de novo lipid A modifications, such as those associated with antimicrobial resistance phenotypes, during Gram-negative bacterial infection and will advance our understanding of the host-pathogen interface.

Original languageEnglish
Pages (from-to)13667-13671
Number of pages5
JournalAnalytical Chemistry
Issue number20
Publication statusPublished - 20 Oct 2020




Dive into the research topics of 'On-Tissue Derivatization of Lipopolysaccharide for Detection of Lipid A Using MALDI-MSI'. Together they form a unique fingerprint.

Cite this