Zhou Xuewen, Zhang Xixi, Boualavong Jonathan, Durney Andrew R, Wang Tonghui, Kirschner Scott, Wentz Michaela, Mukaibo Hitomi
Department of Chemical Engineering, University of Rochester, Rochester, NY, USA.
Electrophoresis. 2017 Oct;38(20):2587-2591. doi: 10.1002/elps.201600548. Epub 2017 May 10.
Electrokinetically controlled microinjection is reported as an effective transport mechanism for microinjection into the wild-type strain of the widely studied model microalga Chlamydomonas reinhardtii. A microinjection system using glass capillary pipettes was developed to capture and impale the motile cells. To apply an electric field and induce electrokinetic flow (e.g., electrophoresis and electroosmosis), an electrode was inserted directly into the solution inside the impaling injection pipette and another electrode was inserted into the external cell media. The viability of the impaled cells was confirmed for more than an hour under 0.01 V using the fluorescein diacetate/propidium iodide dual fluorescent dye based assay. The viability was also found to increase almost logarithmically with decreasing voltage and to depend strongly on the solution within the injection pipette. Successful electrokinetic microinjection into cells was confirmed by both an increase in cell volume under an applied voltage and electric field dependent delivery of fluorescent fluorescein molecules into an impaled cell. Our study offers novel opportunities for quantitative delivery of biomolecules into microalgae and advancing the research and development of these organisms as biosynthetic factories.
据报道，电动控制显微注射是一种有效的运输机制，可用于将其显微注射到广泛研究的模式微藻莱茵衣藻的野生型菌株中。开发了一种使用玻璃毛细管移液器的显微注射系统，用于捕获和刺穿活动细胞。为了施加电场并诱导电动流动（例如，电泳和电渗），将一个电极直接插入刺穿注射移液器内的溶液中，另一个电极插入外部细胞培养基中。使用基于荧光素二乙酸酯/碘化丙啶双荧光染料的检测方法，证实了在0.01 V电压下，被刺穿的细胞在一个多小时内具有活力。还发现活力几乎随着电压的降低呈对数增加，并且强烈依赖于注射移液器内的溶液。通过施加电压下细胞体积的增加以及荧光素分子在电场作用下向被刺穿细胞的递送，证实了成功地对细胞进行了电动显微注射。我们的研究为将生物分子定量递送至微藻以及推动这些生物作为生物合成工厂的研究与开发提供了新的机会。
