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Isolation and enumeration of bacteria at ultralow concentrations and antibiotic resistance profiling are of great importance for early diagnosis and treatment of bacteremia. In this work, we describe a simple, rapid, and versatile magnetic-assisted microfluidic method for rapid bacterial detection. The developed method enables magnetophoretic loading of bead-captured bacteria into the microfluidic chamber under external static and dynamic magnetic fields in 4 min. A shallow microfluidic chamber design that enables the monolayer orientation and transportation of the beads and a glass substrate with a thickness of 0.17 mm was utilized to allow high-resolution fluorescence imaging for quantitative detection. Escherichia coli (E. coli) with green fluorescent protein (GFP)-expressing gene and streptavidin-modified superparamagnetic microbeads were used as model bacteria and capturing beads, respectively. The …