In this paper, we describe a bio-fluidic device for adaptive Sample pretreatment, in order to optimize the conditions under which absorbance assays can be conducted. This device can be successfully applied to the measurement of Escherichia coli (E. coli) concentrations using adaptive dilution, with which the dilution ratio can be adjusted during the dilution. Although many attempts have been previously made to miniaturize complex biochemical analyses at the chip scale, very few sample pretreatment processes have actually been miniaturized or automated at this point. Due to the lack of currently available on-chip pretreatments, analytical instruments tend to suffer from a limited range of analysis. This occasionally hinders the direct and quantitative analysis of specific analytes obtained from real samples. In order to overcome these issues, we exploit two novel strategies: dilution with a programmable ratio, and to-and-fro mixing. The bio-fluidic device consists of a rectangular chamber constructed of poly(dimethylsiloxane) (PDMS). This chamber has four openings, an inlet, an outlet, an air control, and an air vent. Each of the dilution cycles is comprised of four steps: detection, liquid drain, buffer injection, and to-and-fro mixing. When using adaptive sample pretreatment, the range in which E. coli concentrations can be measured is broadened, to an optical density (O.D.) range of 0.3 similar to 30. This device may prove useful in the on-line monitoring of cell concentrations, in both fermenter and aqueous environments.