NEAR INFRARED FLUOROGEN AND FLUORESCENT ACTIVATING PROTEINS FOR IN VIVO IMAGING AND LIVE-CELL BIOSENSING
申请人:Carnegie Mellon University
公开号:US20140243509A1
公开(公告)日:2014-08-28
Tissue slices and whole organisms offer substantial challenges to fluorescence imaging. Autofluorescence and absorption via intrinsic chromophores, such as flavins, melanin, and hemoglobins, confound and degrade output from all fluorescent tags. An “optical window,” farther red than most autofluorescence sources and in a region of low hemoglobin and water absorbance, lies between 650 and 900 nm. This valley of relative optical clarity is an attractive target for fluorescence-based studies within tissues, intact organs, and living organisms. Novel fluorescent tags were developed herein, based upon a genetically targeted fluorogen activating protein and cognate fluorogenic dye that yields emission with a peak at 733 nm exclusively when complexed as a “fluoromodule”. This tool improves substantially over previously described far-red/NIR fluorescent proteins in terms of brightness, wavelength, and flexibility by leveraging the flexibility of synthetic chemistry to produce novel chromophores.
组织切片和整个生物体对荧光成像提出了重大挑战。自发荧光和内在色素(如黄酮,黑色素和血红蛋白)的吸收会干扰和降解所有荧光标记的输出。在650到900纳米之间,比大多数自发荧光源更远的“光学窗口”处于低血红蛋白和水吸收区域内,是组织,完整器官和活体内荧光研究的有吸引力的目标。本文开发了一种新型荧光标记,基于基因定向荧光原活化蛋白和相应的荧光原染料,仅当复合为“荧光模块”时,才产生峰值为733纳米的发射。通过利用合成化学的灵活性来生产新的色素,该工具在亮度,波长和灵活性方面显着优于先前描述的远红外/近红外荧光蛋白。