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Hemifusion of Cells Using Femtosecond Laser Pulses

机译：飞秒激光脉冲半融合细胞

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摘要

Attachment of single cells via hemifusion of cellular membranes using femtosecond laser pulses is reported in this manuscript. This is a method to attach single cells using sub-10 femtosecond laser pulses, with 800 nm central wavelength delivered from a Ti:Sapphire laser is described. A fluorescent dye, Calcein AM, was used to verify that the cell's cytoplasm did not migrate from a dyed cell to a non-dyed cell, in order to ascertain that the cells did not go through cell-fusion process. An optical tweezer was used in order to assess the mechanical integrity of the attached joint membranes. Hemifusion of cellular membranes was successful without initiating full cell fusion. Attachment efficiency of 95% was achieved, while the cells' viability was preserved. The attachment was performed via the delivery of one to two trains of sub-10 femtosecond laser pulses lasting 15 milliseconds each. An ultrafast reversible destabilization of the phospholipid molecules in the cellular membranes was induced due to a laser-induced ionization process. The inner phospholipid cell membrane remained intact during the attachment procedure, and cells' cytoplasm remained isolated from the surrounding medium. The unbounded inner phospholipid molecules bonded to the nearest free phospholipid molecule, forming a joint cellular membrane at the connection point. The cellular membrane hemifusion technique can potentially provide a platform for the creation of engineered tissue and cell cultures.
机译：该手稿报道了使用飞秒激光脉冲通过细胞膜的半融合附着单个细胞。描述了使用低于10飞秒的激光脉冲附着单个细胞的方法，该方法从Ti：蓝宝石激光器发出的中心波长为800 nm。为了确定细胞没有经过细胞融合过程，使用了荧光染料Calcein AM来验证细胞的胞质是否没有从染色的细胞迁移到未染色的细胞。使用光学镊子以评估所附着关节膜的机械完整性。细胞膜的半融合是成功的，没有启动全细胞融合。达到95％的附着效率，同时保留了细胞的活力。通过传递一到两列10毫秒以下的飞秒激光脉冲（每次持续15毫秒）来执行连接。由于激光诱导的电离过程，导致了细胞膜中磷脂分子的超快可逆去稳定作用。在附着过程中，内部磷脂细胞膜保持完好无损，并且细胞质保持与周围介质分离。未结合的内部磷脂分子与最接近的游离磷脂分子结合，在连接点形成关节细胞膜。细胞膜半融合技术可以潜在地为创建工程组织和细胞培养物提供平台。

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  • 来源
    《Optical interactions with tissue and cells XXVI》|2015年|932105.1-932105.7|共7页
  • 会议地点 San Francisco CA(US)
  • 作者

    Nir Katchinskiy; Roseline Godbout; Helly R. Goez; Abdulhakem Y. Elezzabi;

  • 作者单位

    Department of Electrical and Computer Engineering, Ultrafast Photonics and Nano-Optics Laboratory, University of Alberta, Edmonton, Alberta T6G 2V4, Canada;

    Department of Oncology, University of Alberta, Cross Cancer Institute, 11560 University Avenue, Edmonton, Alberta T6G 1Z2, Canada;

    Division of Pediatric Neurology, Department of Pediatrics, Faculty of Medicine and Dentistry, University of Alberta, Alberta T6G 2R7, Canada;

    Department of Electrical and Computer Engineering, Ultrafast Photonics and Nano-Optics Laboratory, University of Alberta, Edmonton, Alberta T6G 2V4, Canada;

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