Single-Cell Chemical Lysis on Microfluidic Chips with Arrays of Microwells 英文参考文献.docVIP

Sensors 2012, 12, 347-358; doi:10.3390/s120100347 OPEN ACCESS sensors ISSN 1424-8220 /journal/sensors Article Single-Cell Chemical Lysis on Microfluidic Chips with Arrays of Microwells Chun-Ping Jen 1,*, Ju-Hsiu Hsiao 1 and Nikolay A. Maslov 2 1 Department of Mechanical Engineering and Advanced Institute of Manufacturing with High-Tech Innovation, National Chung Cheng University, Chia Yi 62102, Taiwan; E-Mail: ggina03@.tw 2 Khristianovich Institute of Theoretical and Applied Mechanics, Siberian Division, Russian Academy of Science, Novosibirsk 630090, Russia; E-Mail: nmaslov@itam.nsc.ru * Author to whom correspondence should be addressed; E-Mail: imecpj@.tw; Tel.: +886-5-272-0411 ext. 33322; Fax: +886-5-272-0589. Received: 29 November 2011; in revised form: 26 December 2011 / Accepted: 29 December 2011 / Published: 30 December 2011 Abstract: Many conventional biochemical assays are performed using populations of cells to determine their quantitative biomolecular profiles. However, population averages do not reflect actual physiological processes in individual cells, which occur either on short time scales or nonsynchronously. Therefore, accurate analysis at the single-cell level has become a highly attractive tool for investigating cellular content. Microfluidic chips with arrays of microwells were developed for single-cell chemical lysis in the present study. The cellular occupancy in 30-μm-diameter microwells (91.45%) was higher than that in 20-μm-diameter microwells (83.19%) at an injection flow rate of 2.8μL/min. However, most of the occupied 20-μm-diameter microwells contained individual cells. The results of chemical lysis experiments at the single-cell level indicate that cell membranes were gradually lysed as the lysis buffer was injected; they were fully lysed after 12 s. Single-cell chemical lysis was demonstrated in the proposed microfluidic chip, which is suitable for high-throughput cell