Katchinskiy Nir, Godbout Roseline, Goez Helly R, Elezzabi Abdulhakem Y
Department of Electrical and Computer Engineering, Ultrafast Photonics and Nano-Optics Laboratory, University of Alberta, Edmonton, AB, T6G 2V4, Canada.
Lasers Surg Med. 2014 Apr;46(4):335-41. doi: 10.1002/lsm.22230. Epub 2014 Feb 18.
Laser-induced cell-cell surgical attachment using femtosecond laser pulses is reported.
STUDY DESIGN/MATERIALS AND METHODS: We have demonstrated the ability to attach single cells using sub-10 femtosecond laser pulses, with 800 nm central wavelength delivered from a Ti:Sapphire laser. To check that the cells did not go through a cell-fusion process, a fluorescent dye Calcein AM was used to verify that the fluorescent dye did not migrate from a dyed cell to a non-dyed cell. The mechanical integrity of the attached joint was assessed using an optical tweezer.
Attachment of cells was performed without the induction of cell-cell fusion, with attachment efficiency of 95%, and while preserving the cells' viability. Cell-cell attachment was achieved by delivery of one to two trains of femtosecond laser pulses lasting 15 ms each.
Laser-induced ionization process led to an ultrafast reversible destabilization of the phospholipid layer of the cellular membrane. The inner cell membrane remained intact during the attachment procedure, and isolation of the cells' cytoplasm from the surrounding medium was obtained. A strong physical attachment between the cells was obtained due to the bonding of the membranes' ionized phospholipid molecules and the formation of a joint cellular membrane at the connection point. The cellular attachment technique, femtosecond laser-induced cell-cell surgical attachment, can potentially provide a platform for the creation of engineered tissue and cell cultures.
报道了使用飞秒激光脉冲诱导细胞间手术附着。
研究设计/材料与方法:我们已证明能够使用中心波长为800nm、由钛宝石激光器发出的亚10飞秒激光脉冲附着单个细胞。为检查细胞未经历细胞融合过程,使用荧光染料钙黄绿素乙酰甲酯来验证荧光染料未从染色细胞迁移至未染色细胞。使用光镊评估附着关节的机械完整性。
细胞附着在不诱导细胞间融合的情况下进行,附着效率为95%,且同时保持细胞活力。通过输送一到两列持续时间各为15毫秒的飞秒激光脉冲实现细胞间附着。
激光诱导的电离过程导致细胞膜磷脂层超快可逆失稳。在附着过程中,细胞内膜保持完整,实现了细胞细胞质与周围介质的隔离。由于膜的电离磷脂分子结合以及在连接点形成联合细胞膜,细胞间获得了牢固的物理附着。细胞附着技术,即飞秒激光诱导细胞间手术附着,有可能为构建工程组织和细胞培养提供一个平台。