期刊
ANALYTICAL CHEMISTRY
卷 80, 期 23, 页码 9141-9149出版社
AMER CHEMICAL SOC
DOI: 10.1021/ac8016423
关键词
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资金
- Core Research for Evolutional Science and Technology (CREST)
- Japan Science and Technology Agency (JST)
- Ministry of Education, Culture, Sports, Science and Technology of Japan
- Human Frontier Science Program
- Kazusa DNA Research Institute
- RIKEN Research Center for Allergy and Immunology
- Toray Science Foundation
Analyzing the intracellular contents and enzymatic activities of single cells is important for studying the physiological and pathological activities at the cellular level. For this purpose, we developed a simple single-cell lysis method by using a dense array of microwells of 10-30-pL volume fabricated by poly(dimethylsiloxane) (PDMS) and a commercially available cell lysis reagent. To demonstrate the performance of this single-cell lysis method, we carried out two different assays at the single-cell level: detection of proteins by antibody conjugated microbeads and measurement of protease activity by fluorescent substrates. The results indicated that this method readily enabled us to monitor protein levels and enzymatic activities in a single cell. Because this method required only an array of PDMS microwells and a fluorescence microscope, the simplicity of this platform opens a way to explore the biochemical characteristics of single cells even by those who are not familiar with microfluidic technology.
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