There have been some studies on the microscopic properties of ester-functionalized ionic liquids (ILs), but the microscopic properties of their mixtures with co-solvents have seldom been reported. In practical applications, ILs are usually used together with co-solvents. Therefore, it is very important to study the microstructure of ester-functionalized ILs and co-solvents. In this work, the hydrogen bonding interactions between ester-functionalized IL 1-acetoxyethyl-3-methylimidazolium tetrafluoroborate (AOEMIMBF4) and DMSO were studied using spectroscopic methods and quantum chemical calculations. Non-functionalized IL 1-butyl-3-methylimidazolium tetrafluoroborate (BMIMBF4) and DMSO were used for comparison. The results indicate that (1) by adding DMSO, the hydrogen bonding interactions of ν(C2-H) were enhanced, and DMSO could form hydrogen bonds with anions and cations simultaneously. (2) The incorporation of an ester group could enhance the hydrogen bonding interactions. (3) Both the stretching vibration of C2-H and CO indicated changes in the microscopic structure: AOEMIMBF4 ion clusters first interacted with DMSO, then broke into AOEMIMBF4-DMSO complexes and finally existed as [AOEMIM]+/[BF4]--DMSO complexes.