title: Fully automated on-chip imaging flow cytometry system with disposable contamination-free plastic re-cultivation chip.
abstract: We have developed a novel imaging cytometry system using a poly(methyl methacrylate (PMMA)) based microfluidic chip. The system was contamination-free, because sample suspensions contacted only with a flammable PMMA chip and no other component of the system. The transparency and low-fluorescence of PMMA was suitable for microscopic imaging of cells flowing through microchannels on the chip. Sample particles flowing through microchannels on the chip were discriminated by an image-recognition unit with a high-speed camera in real time at the rate of 200 event/s, e.g., microparticles 2.5 μm and 3.0 μm in diameter were differentiated with an error rate of less than 2%. Desired cells were separated automatically from other cells by electrophoretic or dielectrophoretic force one by one with a separation efficiency of 90%. Cells in suspension with fluorescent dye were separated using the same kind of microfluidic chip. Sample of 5 μL with 1 × 10(6) particle/mL was processed within 40 min. Separated cells could be cultured on the microfluidic chip without contamination. The whole operation of sample handling was automated using 3D micropipetting system. These results showed that the novel imaging flow cytometry system is practically applicable for biological research and clinical diagnostics.
authors: Kanagawa Academy of Science and Technology, KSP East 310, 3-2-1 Sakado, Takatsu-Ku, Kawasaki 213-0012, Japan; E-Mails: hayashi.bmi@gmail.com (M.H.); hattori.bmi@gmail.com (A.H.); kim.bmi@gmail.com (H.K.); terazono.bmi@gmail.com (H.T.).
authors: Masahito
authors: M
authors: Hayashi
authors: Akihiro
authors: A
authors: Hattori
authors: Hyonchol
authors: H
authors: Kim
authors: Hideyuki
authors: H
authors: Terazono
authors: Tomoyuki
authors: T
authors: Kaneko
authors: Kenji
authors: K
authors: Yasuda
mesh_terms: D000818:Animals; D001331:Automation; D002469:Cell Separation; D005434:Flow Cytometry / Q000295:instrumentation*; D005456:Fluorescent Dyes / Q000737:chemistry; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation; D018715:Microscopy, Video; D032383:Myocytes, Cardiac / Q000166:cytology; D019904:Polymethyl Methacrylate / Q000737:chemistry
chemical_list: D005456:Fluorescent Dyes; D019904:Polymethyl Methacrylate