López-Quesada C, Fontaine A-S, Farré A, Joseph M, Selva J, Egea G, Ludevid M D, Martín-Badosa E, Montes-Usategui M
Optical Trapping Lab - Grup de Biofotònica, Departament de Física Aplicada i Òptica, Universitat de Barcelona, Martí i Franquès 1, 08028 Barcelona, Spain.
Department of Molecular Genetics, Center for Research in Agricultural Genomics (CRAG) CSIC-IRTA-UAB-UB, Campus Universitat Autònoma de Barcelona, Bellaterra (Cerdanyola del Vallès), 08193 Barcelona, Spain.
Biomed Opt Express. 2014 May 30;5(7):1993-2008. doi: 10.1364/BOE.5.001993. eCollection 2014 Jul 1.
Optical trapping supplies information on the structural, kinetic or rheological properties of inner constituents of the cell. However, the application of significant forces to intracellular objects is notoriously difficult due to a combination of factors, such as the small difference between the refractive indices of the target structures and the cytoplasm. Here we discuss the possibility of artificially inducing the formation of spherical organelles in the endoplasmic reticulum, which would contain densely packed engineered proteins, to be used as optimized targets for optical trapping experiments. The high index of refraction and large size of our organelles provide a firm grip for optical trapping and thereby allow us to exert large forces easily within safe irradiation limits. This has clear advantages over alternative probes, such as subcellular organelles or internalized synthetic beads.
光镊可提供有关细胞内部成分的结构、动力学或流变学特性的信息。然而,由于多种因素的综合作用,如目标结构与细胞质的折射率差异较小,对细胞内物体施加显著的力是出了名的困难。在此,我们讨论在内质网中人工诱导形成球形细胞器的可能性,这些细胞器将包含紧密堆积的工程蛋白,用作光镊实验的优化目标。我们的细胞器具有高折射率和大尺寸,为光镊提供了牢固的捕获能力,从而使我们能够在安全的辐照范围内轻松施加较大的力。这相对于其他替代探针,如亚细胞器或内化的合成珠,具有明显优势。