biostudies-literatureMcCaughey CSNational Institute of Allergy and Infectious DiseasesUniversity of California, Santa CruzNIAID NIH HHSDivision of Integrative Organismal SystemsNational Institute of General Medical SciencesNIGMS NIH HHS8308-8316https://www.ebi.ac.uk/biostudies/studies/S-EPMC11140670biostudies-literatureUnknown96(21)Microbial biofilms represent an important lifestyle for bacteria and are dynamic three-dimensional structures. Cyclic dimeric guanosine monophosphate (c-di-GMP) is a ubiquitous signaling molecule that is known to be tightly regulated with biofilm processes. While measurements of global levels of c-di-GMP have proven valuable toward understanding the genetic control of c-di-GMP production, there is a need for tools to observe the local changes of c-di-GMP production in biofilm processes. We have developed a label-free method for the direct detection of c-di-GMP in microbial colony biofilms using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI). We applied this method to the enteric pathogen <i>Vibrio cholerae</i>, the marine symbiont <i>V. fischeri</i>, and Analytical chemistryA Label-Free Approach for Relative Spatial Quantitation of c-di-GMP in Microbial Biofilms.PMC11140670R35GM148385R01 AI102584R01AI10258422205112220510R35 GM148385K12 GM119955Sanchez LMYildiz FHMandel MJMcCaughey CSIsenberg RYTrebino MAMcAtamney AfalseA Label-Free Approach for Relative Spatial Quantitation of c-di-GMP in Microbial Biofilms.Microbial biofilms represent an important lifestyle for bacteria and are dynamic three-dimensional structures. Cyclic dimeric guanosine monophosphate (c-di-GMP) is a ubiquitous signaling molecule that is known to be tightly regulated with biofilm processes. While measurements of global levels of c-di-GMP have proven valuable toward understanding the genetic control of c-di-GMP production, there is a need for tools to observe the local changes of c-di-GMP production in biofilm processes. We have developed a label-free method for the direct detection of c-di-GMP in microbial colony biofilms using matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI). We applied this method to the enteric pathogen <i>Vibrio cholerae</i>, the marine symbiont <i>V. fischeri</i>, and 2024-01-01T00:00:00Z2024 May2026-06-03T03:58:41.004Z2025-08-14T03:05:50.211ZS-EPMC111406703875254310.1021/acs.analchem.3c04687