Department of Biosystems Science and Engineering (D-BSSE), ETH Zurich, Basel, Switzerland.
Department of Molecular and Computational Biology, University of Southern California, Los Angeles, California, USA.
Nat Methods. 2015 Jul;12(7):645-8. doi: 10.1038/nmeth.3405. Epub 2015 May 18.
Spatially confined green-to-red photoconversion of fluorescent proteins with high-power, pulsed laser illumination is negligible, thus precluding optical selection of single cells in vivo. We report primed conversion, in which low-power, dual-wavelength, continuous-wave illumination results in pronounced photoconversion. With a straightforward addition to a conventional confocal microscope, we show confined primed conversion in living zebrafish and reveal the complex anatomy of individual neurons packed between neighboring cells.
利用高功率、脉冲激光照射对荧光蛋白进行空间限制的绿到红光的光致变色转换可以忽略不计,因此无法通过光学手段在体内选择单个细胞。我们报告了引发的转换,其中低功率、双波长、连续波照射会导致明显的光致变色。通过对传统共聚焦显微镜进行简单的改进,我们在活体斑马鱼中实现了受限的引发转换,并揭示了单个神经元在相邻细胞之间的复杂结构。