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Electrophysiological studies of human colonic epithelia traditionally have been hampered by the lack of tissue availability and by poor tissue quality. Human colonic epithelium is usually obtained surgically from individuals with underlying disease, while surgery itself can injure or alter the resected tissue. As a result, a wide range in electrophysiological parameters is reported in previous studies of human colonic epithelium. Such factors may also account for differences in measurements between humans and the few other species studied. We therefore devised a novel and rapid endoscopic technique, endoscopic mucosal resection (EMR), that allows for the removal and study of intestinal mucosal epithelium from normal volunteers. Using EMR we rapidly (7.2 ± 2.4 min) isolated surgical-sized epithelial sheets from the distal colon (1.4 ± 0.4 by 1.3 ± 0.4 cm) that were readily mounted in a 0.64-cm2 Ussing chamber. We observed stable resistance (289 ± 30 Ω · cm2), potential difference (1.6 ± 0.6 mV), and ISC (24 ± 9 μA/cm2) for at least 90 min, after which all experiments were terminated. Exposure to carbachol increased ISC 2.2 ± 0.5-fold, while forskolin increased ISC 4.4 ± 0.5-fold. These data show that the electrophysiological characteristics of the human distal colon removed by EMR more closely approximate values reported for other mammals than when removed using other techniques. Thus EMR represents a significant advance over traditional techniques for isolating human tissues and will increase the availability of this tissue for future studies.