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Rapid Identification of ESKAPE Bacterial Strains Using an Autonomous Microfluidic Device 1 2 3 3 1,3 Jack Y. Ho , Nate J. Cira , John A. Crooks , Josue Baeza , Douglas B. Weibel * 1 Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, Wisconsin, United States of America, 2 Department of Bioengineering, Stanford University, Stanford, California, United States of America, 3 Department of Biochemistry, University of Wisconsin-Madison, Madison, Wisconsin, United States of America Abstract This article describes Bacteria ID Chips (‘BacChips’): an inexpensive, portable, and autonomous microfluidic platform for identifying pathogenic strains of bacteria. BacChips consist of a set of microchambers and channels molded in the elastomeric polymer, poly(dimethylsiloxane) (PDMS). Each microchamber is preloaded with mono-, di-, or trisaccharides and dried. Pressing the layer of PDMS into contact with a glass coverslip forms the device; the footprint of the device in this article is ,6 cm2. After assembly, BacChips are degased under large negative pressure and are stored in vacuum-sealed plastic bags. To use the device, the bag is opened, a sample containing bacteria is introduced at the inlet of the device, and the degased PDMS draws the sample into the central channel and chambers. After the liquid at the inlet is consumed, air is drawn into the BacChip via the inlet and provides a physical barrier that separates the liquid samples in adjacent microchambers. A pH indicator is admixed with the samples prior to their loading, enabling the metabolism of the dissolved saccharides in the microchambers to be