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To use hyperpolarized (HP) 3He MR imaging to assess functional lung ventilation in subjects with cystic fibrosis (CF) before and after treatment.We performed HP 3He static ventilation MRI scans on three subjects, using a Philips 3.0 Tesla (T) Achieva MRI scanner, before and after 11 days of in-patient treatment with combined intravenous and inhaled therapies for pulmonary exacerbations of CF. We also collected spirometry data. We quantified pulmonary ventilation volume measured with HP 3He MRI using an advanced semi-automated analysis technique.Following 11 days of treatment with intravenous antibiotics, hypertonic saline, and rhDNase, HP 3He MR images in one subject displayed a 25% increase in total ventilation volume. Total ventilation volume in the other two subjects slightly decreased. All three subjects showed increases in FEV1 and FVC following treatment.In all subjects, the HP 3He MR images provided detailed information on precisely where in the lungs gas was reaching. These data provide additional support for the conclusion that HP noble gas MRI can be a powerful tool for evaluating lung ventilation in patients with cystic fibrosis, but also raise important questions about the correlation between spirometry and HP gas MRI measurements. J. Magn. Reson. Imaging 2011. © 2011 Wiley Periodicals, Inc.