Software-aided automatic laser optoporation and transfection of cells.
Software-aided automatic laser optoporation and transfection of cells.
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DOI：
10.1038/srep11185
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发表时间：
2015-06-08
影响因子：
4.6
通讯作者：
König K
中科院分区：
文献类型：
--
作者：
Georg Breunig H;Uchugonova A;Batista A;König K
光穿孔，通过激光脉冲使细胞膜透化，已经成为一种强大的非侵入性和高效的诱导细胞转染的技术。然而，通常繁琐的手动靶向单个细胞显著限制了可寻址的细胞数量。为了克服这一限制，我们提出了一个实验装置与定制的软件控制，计算机自动化细胞optoporation。软件评估细胞轮廓的图像对比度，自动指定激光照射的细胞位置，将这些位置置于激光焦点的中心，并执行照射。通过软件控制的样品台的弯曲，原则上可以靶向典型细胞培养皿中的所有细胞，而无需进一步的用户交互。与手动方法相比，自动化允许显著增加可处理细胞的数量。对于100 ms的激光照射持续时间，在显微镜视野区域内每秒可以靶向不同细胞上的7-8个位置。在中国仓鼠卵巢细胞的实验中说明了该装置的实验能力。此外，激光功率的影响进行了讨论，与处理后的细胞存活率和optoporation效率率提到。
Optoporation, the permeabilization of a cell membrane by laser pulses, has emerged as a powerful non-invasive and highly efficient technique to induce transfection of cells. However, the usual tedious manual targeting of individual cells significantly limits the addressable cell number. To overcome this limitation, we present an experimental setup with custom-made software control, for computer-automated cell optoporation. The software evaluates the image contrast of cell contours, automatically designates cell locations for laser illumination, centres those locations in the laser focus, and executes the illumination. By software-controlled meandering of the sample stage, in principle all cells in a typical cell culture dish can be targeted without further user interaction. The automation allows for a significant increase in the number of treatable cells compared to a manual approach. For a laser illumination duration of 100 ms, 7-8 positions on different cells can be targeted every second inside the area of the microscope field of view. The experimental capabilities of the setup are illustrated in experiments with Chinese hamster ovary cells. Furthermore, the influence of laser power is discussed, with mention on post-treatment cell survival and optoporation-efficiency rates.
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影响因子：
4.3
作者：
Kim, Tae Kyung;Eberwine, James H.
通讯作者：
Eberwine, James H.
影响因子：
3.8
作者：
Uchugonova, Aisada;Koenig, Karsten;Tempea, Gabriel
通讯作者：
Tempea, Gabriel
影响因子：
1.6
作者：
KAPUSCINSKI, J
通讯作者：
KAPUSCINSKI, J
影响因子：
3.2
作者：
Krishan, A;Dandekar, PD
通讯作者：
Dandekar, PD
影响因子：
120.1
作者：
Ebert, Allison D.;Svendsen, Clive N.
通讯作者：
Svendsen, Clive N.