Fe-TAMLs as a new class of small molecule peroxidase probes for correlated light and electron microscopy.
Fe-TAMLs as a new class of small molecule peroxidase probes for correlated light and electron microscopy.
复制标题
Fe-TAML 作为一类新型小分子过氧化物酶探针，用于相关光学和电子显微镜。
DOI：
10.1101/2023.08.25.554352
复制
发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Ellisman,MarkH
中科院分区：
文献类型：
--
作者：
Adams,StephenR;Mackey,MasonR;Ramachandra,Ranjan;Deerinck,ThomasJ;Castillon,GuillaumeA;Phan,Sebastien;Hu,Junru;Boassa,Daniela;Ngo,JohnT;Ellisman,MarkH
我们介绍Fe-TAML，一种基于小分子的过氧化物酶，作为相关荧光和电子显微镜工具箱的多功能新成员。探针的实用性通过对新合成的DNA（通过双正交标记）、基因标记的蛋白质（使用HaloTag）和未标记的内源性蛋白质（通过免疫染色）的高分辨率成像得到了证明。利用Fe-TAML催化沉积基于聚合3,3 ' -二氨基苯胺的局部亲锇沉淀物的活性，促进了这些应用中的EM可视化。本文还介绍了Fe-TAML探针合成和实现的优化条件。总之，Fe-TAML是一种新的化学生物学工具，可用于在纳米和微米尺度上可视化细胞内不同的生物分子物种。
We introduce Fe-TAML, a small molecule-based peroxidase as a versatile new member of the correlated fluorescence and electron microscopy toolkit. The utility of the probe is demonstrated by high resolution imaging of newly synthesized DNA (through biorthogonal labeling), genetically tagged proteins (using HaloTag), and untagged endogenous proteins (via immunostaining). EM visualization in these applications is facilitated by exploiting Fe-TAML’s catalytic activity for the deposition of localized osmiophilic precipitates based on polymerized 3,3’-diaminobenzidine. Optimized conditions for synthesizing and implementing Fe-TAML based probes are also described. Overall, Fe-TAML is a new chemical biology tool that can be used to visualize diverse biomolecular species along nanometer and micron scales within cells.