We introduce flexible microfluidic device to characterize the mechanical properties of soft viscoelastic solids such as bacterial biofilms. In the device, stress is imposed on a test specimen by the application of a fixed pressure to a thin, flexible poly(dimethyl siloxane) (PDMS) membrane that is in contact with the specimen. The stress is applied by pressurizing a microfabricated air channel located above the test area. The strain resulting from the applied stress is quantified by measuring the membrane deflection with a confocal laser scanning microscope. The deflection is governed by the viscoelastic properties of the PDMS membrane and of the test specimen. The relative contributions of the membrane and test material to the measured deformation are quantified by comparing a Finite clement analysis with an independent (control) measurement of the PDMS membrane mechanical properties. The flexible microfluidic rheometer was used to characterize both the steady-state elastic modulus and the transient strain recoil of two soft materials: gellan gums and bacterial biofilms. The measured linear elastic moduli and viscoelastic relaxation times of gellan gum solutions were in good agreement with the results of conventional mechanical rheometry. The linear Young's moduli of biofilms of Staphylococcus epidermidis and Klebsiella pneumoniae, which could not be measured using conventional Methods, were found to be 3.2 and 1.1 kPa, respectively, and the relaxation time of the S. epidermidis biofilm was 13.8 s. Additionally, strain hardening was observed in all the biofilms Studied. Finally, design parameters and detection limits of the method show that the device is capable of characterizing soft viscoelastic solids with elastic moduli in the range of 10(2)-10(5) Pa. The flexible microfluidic rheometer addresses the need for mechanical property characterization of soft viscoelastic solids common in fields such as biomaterials, food, and consumer products. It requires only similar to 200 pL of the test specimen.