MRNA ANALYSIS OF MULTICELLULAR SPHEROID WITH A SCANNING PROBE MICROSCOPY SYSTEM

摘要

Here, we introduce several applications for singlecell gene-expression analysis utilizing electrical cell lysis technique with a Pt-ring electrode probe. Glass capillary probes featuring a Pt-ring electrode were utilized to collect single cell with two or three dimensional culture systems. An electric pulse with 100 V height and 10 μs width was applied to lyse the cellular membrane of the single cell in 0. 2 M sucrose solution. Multicellular spheroid of human breast cancer cell line MCF-7 was formed according to hanging drop method or on-top culture on a matrigel sheet. It was shown that evaluation of mRNA expression level is possible at single-cell level collected at the surface of the multicellular spheroid.