Greulich Karl Otto
Leibniz Institute for Age Research/Fritz Lipmann Institute, D-07745 Jena, Germany.
Methods Cell Biol. 2007;82:59-80. doi: 10.1016/S0091-679X(06)82002-9.
This contribution bridges the gap from early European contributions via laser micromanipulation to recent work on the use of laser microbeams and optical tweezers in studies of basic aspects of heart infarction. Laser transfection, particularly of plant cells and their chloroplasts, and laser microdissection of chromosomes with subsequent generation of chromosome segment-specific DNA libraries and laser-induced cell fusion are reported. With optical tweezers, microgravity can be simulated in roots of the alga Chara. Surprisingly, microgravity reduces growth. In some plant cells, CW lasers, in principle suited primarily for optical tweezers, can be used as microbeam. Also, it is shown that natural killer cells mount an attack on leukemia cells even in the absence of specificity, just induced by exerting force with optical tweezers. Finally, with the help of a laser microbeam, lesions can be induced to study wound healing after heart infarction. A modification of optical tweezers, the erythrocyte-mediated force application (EMFA) technique can be used to induce calcium waves not only in tissue reconstituted from excitable heart muscle cells but also from nonexcitable fibroblasts.
本论文从早期欧洲通过激光显微操作的贡献,到近期在心肌梗死基础研究中使用激光微束和光镊的工作,填补了其间的空白。文中报道了激光转染,特别是植物细胞及其叶绿体的转染,以及染色体的激光显微切割及随后生成染色体片段特异性DNA文库和激光诱导的细胞融合。利用光镊,可以在轮藻的根中模拟微重力。令人惊讶的是,微重力会抑制生长。在一些植物细胞中,原则上主要适用于光镊的连续波激光器可被用作微束。此外,研究表明,即使没有特异性,自然杀伤细胞也会在光镊施加力的作用下对白血病细胞发起攻击。最后,借助激光微束,可以诱导损伤来研究心肌梗死后的伤口愈合。光镊的一种改进形式,即红细胞介导的力施加(EMFA)技术,不仅可用于在由可兴奋心肌细胞重构的组织中诱导钙波,也可用于在不可兴奋的成纤维细胞中诱导钙波。