An Integrated Microfluidic Chip for Single Cells Positioning and Interaction Monitoring

来源 :第八届全国微全分析系统学术会议、第三届全国微纳尺度生物分离分析学术会议暨第五届国际微化学与微系统学术会议 | 被引量 : 0次 | 上传用户:fangfang_936
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  We present a simple technique to real-time screening two single-cells signal interactions on our unique microfluidic chip,which is potentially useful for single-cell-based research.The interactions between cells play essential roles in cell biology.Nowadays reports showed that studying cells to cells interactions is an important research to understand cell behaviors such as variation or tumor progression and metastasis [1],which cannot be fully revealed by traditional methods based on the ensemble average behaviors of large population of cells incubated in petri dish.Thus single cell positioning,incubation and observation have become an important technique to analyze single cell behaviors.In this study,we proposed an unique chip combining micro snare [2] to capture single cells and controllable valves [3] to manipulate fluid operation,for facilitating the study of single cell-cell interactions.In this research,a process including six procedures was presented for studying single-cell interactions.This process can be accomplished by a system consisting mechanical traps and valves as shown in Fig.1.Firstly,two single-cells were captured independently by the 15μmx15μm snare which was designed as the size of the cell.And,the two single-cells underwent perfusion culture.Afterward,cell A was stimulated and released some substance.Then the substance released by cell A stimulates cell B by opening the path between them.Finally,the substance released by cell B passed into the detection part to examine the variation of cell B.Fig.2 illustrates the fabrication procedures to create the chip consisting micro-valves and flow channel structure.The valves could be manipulated by a vacuum pump controlled via Lab-view software.The operating method and testing of the valves is showed in Fig.3.In Fig.4,certain fluid was injected into two different directions to ensure the valves could switch the pathway between two single-cell culturing systems.In this way,the two single-cells could be cultured under independent medium route avoiding mutual mixture and could also be affected by each other by opening the valve between two systems.The process of single cell trapping is demonstrated in Fig.5.After single cell trapped inside the snare,the streamline will not go through the snare center since the increase of the flow resistance and the rest of cells will pass by without accumulation inside the trap.Hela cells dyed with Calcein AM Fluorescent were loaded into the chip and the single-cell was captured by the trap as in Fig.6.We could further use this special design to undergo cell-to-cell interactions monitoring in the future.It is a prospective platform for cell-cell interaction research,which can also combine with biological or electrochemical detection.
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