Fluorescent and Bioluminescent Calcium Indicators with Tuneable Colors and Affinities.

Fluorescent and Bioluminescent Calcium Indicators with Tuneable Colors and Affinities.
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具有可调颜色和亲和力的荧光和生物发光钙指示剂。

DOI:
10.1021/jacs.2c01465
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发表时间:
2022-04-20
影响因子:
15
通讯作者:
Johnsson K
Johnsson K
中科院分区:
化学1区
文献类型:
--
作者:
Mertes N;Busch M;Huppertz MC;Hacker CN;Wilhelm J;Gürth CM;Kühn S;Hiblot J;Koch B;Johnsson K

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我们推出一系列明亮的罗丹明钙指示剂,具有可调的亲和力和颜色。指示剂可以特异性地定位于不同的细胞区室,并且通过与HaloTag融合蛋白缀合而与荧光和生物发光读数相容。重要的是,它们在定位时荧光的增加使得无需洗涤的活细胞成像成为可能,这极大地促进了它们在生物测定中的使用。在大鼠海马神经元中作为荧光指示剂的应用包括单个动作电位和内质网中钙流的检测。作为生物发光指示剂的应用包括在高通量相容测定中记录核钙的药理学调节。这些指示剂的多功能性和显著的易用性使其成为生物成像和生物测定的有力工具。
We introduce a family of bright, rhodamine-based calcium indicators with tuneable affinities and colors. The indicators can be specifically localized to different cellular compartments and are compatible with both fluorescence and bioluminescence readouts through conjugation to HaloTag fusion proteins. Importantly, their increase in fluorescence upon localization enables no-wash live-cell imaging, which greatly facilitates their use in biological assays. Applications as fluorescent indicators in rat hippocampal neurons include the detection of single action potentials and of calcium fluxes in the endoplasmic reticulum. Applications as bioluminescent indicators include the recording of the pharmacological modulation of nuclear calcium in high-throughput compatible assays. The versatility and remarkable ease of use of these indicators make them powerful tools for bioimaging and bioassays.
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发表时间: 2015
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