可用于双色双光子显微成像的新的荧光蛋白对(英文)

A Novel Fluorescent Protein Pair for Dual-color Two-photon Laser Scanning Microscopy

双色双光子激光扫描显微技术可以用来研究生物组织内两种不同蛋白质的表达、定位和示踪.由于大多数双光子显微镜一次只能提供一种波长的激发光,双色同时成像较难实现.mAmetrine和mKate2作为新发现的荧光蛋白对可以用于双光子双色同时成像,这得益于它们各自的优势:mAmetrine的斯托克斯位移和mKate2的高亮度.在765nm的波长激发时,它们的双光子吸收效率都很高.mAmetrine和mKate2能够很好地用于双色双光子活细胞成像实验.

Dual-color two-photon laser scanning microscopy is a useful method for simultaneously studying the expression, localization and trafficking of two different proteins in tissues. Because most two-photon microscopes only use a single wavelength excitation laser, simultaneously exciting multiple fluorescent proteins remains a challenge. Here, we present mAmetrine and mKate2, which can be used as a novel fluorescent protein pair in dual-color two-photon imaging by taking advantage of the large Stokes shift of mAmetrine and high brightness of mKate2. Both proteins have high two-photon absorption efficiencies and can be simultaneously excited at an optical wavelength of 765 nm. Dual-color two-photon imaging using this protein pair is highly effective in living cells.