Dilution is a standard fluid operation widely employed in the sample preparation process of many bio(chemical) assays. It serves multiple essential functions such as sample mixing with certain reagents at specific dilution ratios, reducing sample matrix effects, bringing target analytes within the linear assay detection range, among many others. Traditionally, sample processing is performed in laboratory settings through manual or automated pipetting. When working in resource-limited settings, however, neither trained personnel nor proper laboratory equipment are available limiting the accessibility to high-quality diagnostic tests. In this work, we present a novel standalone and fully automated microfluidic platform for the stepwise preparation of serial dilutions without the need for any active elements. Stepwise dilution is achieved using the coordinated burst action of hydrophobic burst valves to first isolate a precisely metered volume from an applied sample drop and subsequently merge it with a prefilled diluent liquid. Downstream, expansion chambers are used to mix both reagents into a homogeneous solution. The dilution module was characterized to generate accurate and reproducible (CV < 7%) dilutions for targeted dilution factors of 2, 5 and 10×, respectively. Three dilution modules were coupled in series to generate three-fold logarithmic (log₅ or log₁₀) dilutions, with excellent linearity (R² > 0.99). Its compatibility with whole blood was furthermore illustrated, proving its applicability for automating and downscaling bioassays with complex biological matrices. Finally, autonomous on-chip serial dilution was demonstrated by incorporating the self-powered (i)SIMPLE technology as a passive driving source for liquid manipulation. We believe that the simplicity and modularity of the presented autonomous dilution platform are of interest to many point-of-care applications in which sample dilution and reagent mixing are of importance.