Selective removal of ovarian cancer cells from human ascites fluid using magnetic nanoparticles

(A) Fluorescence microscopy image (20×) of superparamagnetic nanoparticles coated with glucuronic acid and Rhodamine-conjugated peptides taken on an Olympus X51, equipped with an Olympus DP-71 camera, and using an Olympus Rhodamine filter. The particles in this image were aggregated using a 2500-gauss magnet to enhance visibility. (B) Microscope bright-field image (10×) of filtrand from ascites sample stained with Trypan Blue. Ascites samples had been formerly stored in 10% dimethyl sulfoxide at –80°C and would contain dead cells that would not prevent Trypan Blue from traversing their membranes. In this image we see the aggregation of dead cells bound to nanoparticles magnetically extracted from the patient samples.

(A–C) Dot plots of flow cytometry analysis of ascites samples for patient 914 (trials 1–3) showing gated populations, frequencies, and population labels (P1–P4). (D–F) Dot plots of flow cytometry analysis of filtrand samples for patient 914 (trials 1–3) showing gates copied from untreated ascites trial (trial 1), frequencies, and population labels (P1–P4). (G–I) Filtrand extracted from patient samples (patient 914) using nanoparticles having no peptide conjugates. Population gates were copied from previous trials performed on untreated ascites and pure nanoparticle samples. (J,K) Dot plots of superparamagnetic nanoparticles coated with glucuronic acid with (J) and without (K) peptide functions showing gates copied from untreated ascites trial (trial 1).

(A–D) Patient sample 914 was stained with a panel of fluorescein isothiocyanate (FITC)-conjugated monoclonal antibodies. The cell counts and fluorescence intensities for cells testing positive in P1–P4 are compared to autofluorescence levels in a sample of unstained ascites. (E–H) Patient sample 914 was stained with phycoerythrin-conjugated antibody (PE-A) to EphA2. The cell counts and fluorescence intensities for cells testing positive in P1–P4 are compared to autofluorescence levels in a sample of unstained ascites.

(A–D) Patient sample 914 was stained with a panel of fluorescein isothiocyanate (FITC)-conjugated monoclonal antibodies. The cell counts and fluorescence intensities for cells testing positive in P1–P4 are compared to autofluorescence levels in a sample of unstained ascites. (E–H) Patient sample 914 was stained with phycoerythrin-conjugated antibody (PE-A) to EphA2. The cell counts and fluorescence intensities for cells testing positive in P1–P4 are compared to autofluorescence levels in a sample of unstained ascites.