High-speed imaging of gas-bubble formation during femtosecond-laser cell optoporation
High-speed imaging of gas-bubble formation during femtosecond-laser cell optoporation
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DOI：
10.1117/12.2542313
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发表时间：
2020-02
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影响因子：
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通讯作者：
A. Batista;H. Breunig;K. König
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文献类型：
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作者：
A. Batista;H. Breunig;K. König
飞秒激光脉冲可以在细胞膜上产生短暂的洞，使其短暂地渗透到遗传大分子中。这是一种非常有效的细胞转染和重编程方法。对于足够高的辐照度值，激光辐射通过多光子电离导致等离子体形成，并且随后形成在照射后不久可见的充气气泡。虽然这种气泡的形成是众所周知的，但基本的微观过程、最佳气泡尺寸和指示瞬时孔形成的持续时间不太清楚。气泡形成和成功的optoporation之间的对应关系进一步复杂的事实，即形成很大程度上取决于照射的细胞位置和激光照射参数（功率，曝光时间）。我们已经研究了气泡的形成，从短脉冲照射两个商业钛：蓝宝石激光器使用高速摄像机。更高的激光功率和更长的曝光时间产生更大的气泡，需要更长的时间才能崩溃。此外，气泡的特性与细胞的代谢和光致发光后的活力之间的相关性被发现，这些结果可以帮助优化激光参数，有效的光致发光和高的后处理细胞的活力，以及照射光的气泡形成背后的微观激发过程。
Femtosecond-laser pulses can create transient holes in the membrane of a cell, making it briefly permeable to genetic macromolecules. This is a highly effective method for cell transfection and reprogramming. For sufficiently high irradiance values, the laser radiation leads to plasma formation through multiphoton ionization and a subsequent formation of gas-filled bubbles which are shortly visible after the irradiation. While this bubble formation is well known, the underlying microscopic processes, the optimal bubble size and duration which indicate the transient hole formation are less clear. The correspondence between bubble formation and successful optoporation is further complicated by the fact that the formation greatly depends on the irradiated cell position and laser irradiation parameters (power, exposure time). We have investigated the formation of bubbles resulting from short pulse irradiation with two commercial Ti:sapphire lasers using a high-speed camera. Higher laser powers and longer exposure times yielded bigger bubbles which took longer to collapse. Additionally, a correlation between the bubble characteristics and the cell’s metabolism and post-optoporation viability was found. These results can help to optimize the laser parameters for efficient optoporation and high post-treatment cell viability as well as to shine light on the microscopic excitation processes behind the bubble formation.