Virus enrichment for single virus infection by using 3D insulator based dielectrophoresis

• Published in: PloS one Vol. 9; no. 2; p. e94083
• Main Authors: Masuda, Taisuke, Maruyama, Hisataka, Honda, Ayae, Arai, Fumihito
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 11.06.2014; Public Library of Science (PLoS)
• Subjects: Applied physics; Bacteria; Biology and Life Sciences; Cell cycle; Cell Line; Dielectrophoresis; Electric fields; Electrodes; Electrophoresis, Microchip - instrumentation; Engineering and Technology; Engineering schools; Enrichment; Equipment Design; Fabrication; Filtration - instrumentation; Flow channels; Genetic engineering; Glass substrates; Health aspects; Humans; Infection; Infections; Influenza; Influenza, Human - virology; Integrated circuit fabrication; Lasers; Lung - cytology; Lung - virology; Medicine and Health Sciences; Microfluidics; Optical Tweezers; Optics; Orthomyxoviridae - isolation & purification; Orthomyxoviridae - physiology; Photolithography; Physiology; Sensors; Substrate inhibition; Three dimensional flow; Virus diseases; Virus Internalization; Viruses
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0094083

We developed an active virus filter (AVF) that enables virus enrichment for single virus infection, by using insulator-based dielectrophoresis (iDEP). A 3D-constricted flow channel design enabled the production of an iDEP force in the microfluidic chip. iDEP using a chip with multiple active virus filters (AVFs) was more accurate and faster than using a chip with a single AVF, and improved the efficiency of virus trapping. We utilized maskless photolithography to achieve the precise 3D gray-scale exposure required for fabrication of constricted flow channel. Influenza virus (A PR/8) was enriched by a negative DEP force when sinusoidal wave was applied to the electrodes within an amplitude range of 20 Vp-p and a frequency of 10 MHz. AVF-mediated virus enrichment can be repeated simply by turning the current ON or OFF. Furthermore, the negative AVF can inhibit virus adhesion onto the glass substrate. We then trapped and transported one of the enriched viruses by using optical tweezers. This microfluidic chip facilitated the effective transport of a single virus from AVFs towards the cell-containing chamber without crossing an electrode. We successfully transported the virus to the cell chamber (v = 10 µm/s) and brought it infected with a selected single H292 cell.