Braschler Thomas, Johann Robert, Heule Martin, Metref Lynda, Renaud Philippe
Microsystems Laboratory, STI-LMIS, Swiss Federal Institute of Technology Lausanne, EPFL, CH-1015 Lausanne, Switzerland.
Lab Chip. 2005 May;5(5):553-9. doi: 10.1039/b417604a. Epub 2005 Mar 15.
Microfluidic devices are increasingly used to perform biological experiments on a single-cell basis. However, long-term stability of cell positions is still an issue. A novel biocompatible method for cell entrapment and release on a microchip is presented. It is based on the controlled formation of an alginate hydrogel by bringing two laminar flows of alginate and calcium ions in the range of 2 mM to 40 mM into contact. The resulting growth of a gel bar is used to enclose and immobilize yeast cells. Adding ethylenediaminetetraacetic acid (EDTA) to the alginate solution allows for control of the hydrogel growth, and by varying the ratio of Ca(2+) to EDTA concentrations gel growth or gel shrinkage can be induced at will. Trapped cells are released during shrinkage of the gel. The trapping efficiency for different cell speeds is investigated and the properties of gel growth are discussed using a diffusion model. Precise positioning of a single cell is demonstrated. The technique presented allows not only the reversible immobilization of cells under gentle conditions but also offers the potential of long-term cell cultures as shown by on-chip incubation of yeast cells. The procedure may provide a simple and fully biocompatible technique for a multitude of innovative experiments on cells in microsystems.
微流控装置越来越多地用于在单细胞基础上进行生物学实验。然而,细胞位置的长期稳定性仍然是一个问题。本文提出了一种在微芯片上捕获和释放细胞的新型生物相容性方法。它基于通过使浓度在2 mM至40 mM范围内的藻酸盐和钙离子的两个层流接触来控制藻酸盐水凝胶的形成。所得凝胶条的生长用于包围和固定酵母细胞。向藻酸盐溶液中添加乙二胺四乙酸(EDTA)可控制水凝胶的生长,并且通过改变Ca(2+)与EDTA浓度的比例,可以随意诱导凝胶生长或凝胶收缩。被困细胞在凝胶收缩期间释放。研究了不同细胞速度下的捕获效率,并使用扩散模型讨论了凝胶生长的特性。展示了单个细胞的精确定位。所提出的技术不仅允许在温和条件下对细胞进行可逆固定,而且还提供了长期细胞培养的潜力,如酵母细胞在芯片上的孵育所示。该方法可为微系统中细胞的大量创新实验提供一种简单且完全生物相容的技术。