Accurate quantitative information of the analytes in mass spectrometry imaging (MSI) is fundamental for determining the accurate spatial distribution, which can provide additional insight into the living processes, disease progression or the pharmacokinetic-pharmacodynamic mechanisms. However, performing a quantitative analysis in MSI is still challenging. This review focuses on the quantitation-related factors and recent advances in the strategies of quantitative MSI (q-MSI) of small molecules. The main quantitation-related factors are discussed according to the new investigations in recent years, including the regionally varied extraction efficiencies and ionization efficiencies, signal-concentration regression functions, and the repeatability of surface sampling/ionization methods. Newly developed quantitation strategies in MSI based on aforementioned factors are introduced, including new techniques in standard curve calibration with normalization to an internal standard, kinetic calibration, and chemometric methods. Different strategies for validating q-MSI methods are discussed. Finally, the future perspectives to q-MSI are proposed.