Organic dyes generally suffer from small Stokes shift that usually leads to self‐quenching and ‐gaining errors during the fluorescent imaging process. Here, a through‐bond energy transfer (TBET) cassette is developed with large Stokes shift to pursue precise cell imaging. The TBET is constructed by covalently conjugated tetraphenylethene (acts as donor) and rhodamine (acceptor) through an acetylene bond. The constructed TBET cassette distinctly behaves as dual‐Stokes shifts, including a large pseudo‐Stokes shift caused by energy transfer, from donor's absorption to acceptor's emission (up to 260 nm) and a smaller Stokes shift of acceptor molecules itself. Due to the intrinsic dual‐Stokes shifts, TBET cassette exhibits specific “dual distinct absorbances, single shared emission” properties, which can be excitated under two different laser channels. By colocalization of the imaging readouts of these two channels, the precisely “double checked” fluorescent imaging is achieved in living cells.

中文翻译：

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具有双斯托克斯位移的贯通键能量转移盒，用于“双重检查”细胞成像

有机染料通常会遭受较小的斯托克斯位移，这通常会导致荧光成像过程中的自猝灭和增益误差。在这里，开发了一种具有大斯托克斯位移的键合能量转移（TBET）盒，以追求精确的细胞成像。TBET 由四苯乙烯（作为供体）和罗丹明（受体）通过乙炔键共价结合而成。构建的TBET盒明显表现为双斯托克斯位移，包括由能量转移引起的大的伪斯托克斯位移，从供体吸收到受体发射（高达260 nm）和受体分子本身的较小斯托克斯位移。由于固有的双斯托克斯位移，TBET盒表现出特定的“双不同吸光度，单共享发射”特性，可以在两个不同的激光通道下激发。通过这两个通道的成像读数的共定位，在活细胞中实现了精确的“双重检查”荧光成像。