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首页> 外文会议>Optical Trapping and Optical Micromanipulation II >Optical trapping inside living organisms
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Optical trapping inside living organisms

机译：生物体内的光阱

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We use optical tweezers to investigate processes happening inside living cells. In a previous study, we trapped naturally occurring lipid granules inside living yeast cells, and used them to probe the viscoelastic properties of the cytoplasm. However, we prefer to use probes which can be specifically attached to various organelles within the living cells in order to optically quantify the forces acting on these organelles. Therefore, we have chosen to use nanometer sized gold beads as probes. These gold beads can be conjugated and attached chemically to the organelles of interest. Only Rayleigh metallic particles can be optically trapped and for these it is the case that the larger the beads, the larger the forces which can be exerted and thus measured using optical tweezers. The gold nanoparticles are injected into the cytoplasm using micropipettes. The very rigid cell wall of the S. pombe yeast cells poses a serious obstacle to this injection. In order to be able to punch a hole in the cell, first, the cells have to be turned into protoplasts, where only a lipid bilayer separates the cytoplasm from the surrounding media. We show how to perform micropipette delivery into the protoplasts and also how the protoplasts can be ablated using the trapping laserlight. Finally, we demonstrate that we can transform the protoplasts back to normal yeast cells.
机译：我们使用光镊来研究活细胞内部发生的过程。在先前的研究中，我们将天然存在的脂质颗粒捕获在活酵母细胞内，并用它们来探测细胞质的粘弹性质。但是，我们更喜欢使用可以特异性附着在活细胞内各种细胞器上的探针，以便从光学角度量化作用在这些细胞器上的力。因此，我们选择使用纳米尺寸的金珠作为探针。这些金珠可以被缀合并化学附接到感兴趣的细胞器。仅瑞利金属颗粒可以被光学捕获，并且对于这些情况，珠子越大，可以施加的力越大，因此可以使用光镊测量。使用微量移液器将金纳米颗粒注入细胞质。粟酒裂殖酵母酵母细胞非常坚硬的细胞壁严重阻碍了这种注射。为了能够在细胞中打孔，首先，必须将细胞转变为原生质体，其中只有脂质双层将细胞质与周围培养基分开。我们展示了如何执行微量移液器传递到原生质体，以及如何使用捕获激光烧蚀原生质体。最后，我们证明了我们可以将原生质体转化回正常的酵母细胞。

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