Versatile system for single cell acquisition and analysis

Abstract

Abstract Single cell analysis (SCA) has become an essential part of basic and clinical research. A prerequisite to SCA studies is the isolation of individual cells from cell cultures. There are commercially available technologies including laser based systems and cell sorting instruments. However, they are typically complex and expensive making integration within standard lab/clinical sample processing workflows difficult. We have developed a universal platform for rapid acquisition of single cells from cultures and deposition into the single wells for further molecular analysis or clonal expansion. The instrument may also be used for tissue microdissection and measurements of individual cell adhesion. The main working principle is based on the capillary based vacuum pulse assisted technology (CTAS; UP8797644). Developed instrument collects individual cells from any adherent cultures grown in standard cell culture dishes in as small as 15 nl volume. It may be used with a wide range of inverted microscopes, so that the cells of interest can be identified based on morphology, location or fluorescent label. Automatic extraction and deposition algorithms were developed for acquisition of targeted cells and tissue regions of interest. Here, individual cells were collected from human neuroblastoma SH-SY5Y, CHO, and 3T3 cell lines. Collected cells were dispensed immediately into individual wells for clonal expansion. Clonal expansion and trypan blue assay revealed minimal effect on cellular viability (up to 99% when compared to controls). The benefits of the proposed technology include cost-efficiency, simple operation, complete workflow, and compatibility with a wide range of inverted microscopes and standard culture dishes.