Fluorochromized tyramide-glucose oxidase as a multiplex fluorescent tyramide signal amplification system for histochemical analysis.
Fluorochromized tyramide-glucose oxidase as a multiplex fluorescent tyramide signal amplification system for histochemical analysis.
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DOI：
10.1038/s41598-022-19085-9
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发表时间：
2022-09-12
影响因子：
4.6
通讯作者：
Hioki, Hiroyuki
中科院分区：
文献类型：
--
作者：
Yamauchi, Kenta;Okamoto, Shinichiro;Ishida, Yoko;Konno, Kohtarou;Hoshino, Kisara;Furuta, Takahiro;Takahashi, Megumu;Koike, Masato;Isa, Kaoru;Watanabe, Masahiko;Isa, Tadashi;Hioki, Hiroyuki
酪胺信号放大(TSA)是一种高度敏感的组织化学分析方法。之前，我们报道了一种TSA系统，生物素酪胺-葡萄糖氧化酶(BT-GO)，用于亮场成像。在这里，我们开发了荧光酪胺-葡萄糖氧化酶(FT-GO)作为一种多重荧光TSA系统。FT-GO是指在过氧化物酶的催化下，葡萄糖和葡萄糖氧化酶反应生成过氧化氢，生成含氟酪胺(FT)的过程。我们发现FT-GO增强了免疫荧光信号，同时保持了低背景信号。与间接免疫荧光法相比，FT-GO法显示单胺能投射系统在小鼠和恒河猴脑中的分布更为广泛。对于FT-GO的多重标记，我们使用叠氮化钠猝灭抗体标记的过氧化物酶。我们将FT-GO应用于多重荧光原位杂交，并结合免疫荧光成功地标记了新皮质中间神经元亚型。FT-GO免疫荧光进一步提高了腺相关病毒示踪剂的检测能力。由于FT-GO的简单性和高信噪比的染色，将为组织化学分析提供一个通用的平台。
Tyramide signal amplification (TSA) is a highly sensitive method for histochemical analysis. Previously, we reported a TSA system, biotinyl tyramine-glucose oxidase (BT-GO), for bright-filed imaging. Here, we develop fluorochromized tyramide-glucose oxidase (FT-GO) as a multiplex fluorescent TSA system. FT-GO involves peroxidase-catalyzed deposition of fluorochromized tyramide (FT) with hydrogen peroxide produced by enzymatic reaction between glucose and glucose oxidase. We showed that FT-GO enhanced immunofluorescence signals while maintaining low background signals. Compared with indirect immunofluorescence detections, FT-GO demonstrated a more widespread distribution of monoaminergic projection systems in mouse and marmoset brains. For multiplex labeling with FT-GO, we quenched antibody-conjugated peroxidase using sodium azide. We applied FT-GO to multiplex fluorescent in situ hybridization, and succeeded in labeling neocortical interneuron subtypes by coupling with immunofluorescence. FT-GO immunofluorescence further increased the detectability of an adeno-associated virus tracer. Given its simplicity and a staining with a high signal-to-noise ratio, FT-GO would provide a versatile platform for histochemical analysis.
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影响因子：
--
作者：
King RS;Newmark PA
通讯作者：
Newmark PA
影响因子：
4.2
作者：
Bockstael, Olivier;Melas, Catherine;Tenenbaum, Liliane
通讯作者：
Tenenbaum, Liliane
影响因子：
5.8
作者：
Furuta T;Yamauchi K;Okamoto S;Takahashi M;Kakuta S;Ishida Y;Takenaka A;Yoshida A;Uchiyama Y;Koike M;Isa K;Isa T;Hioki H
通讯作者：
Hioki H
DOI：
10.1523/jneurosci.0154-12.2012
发表时间：
2012-06-06
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Igarashi KM;Ieki N;An M;Yamaguchi Y;Nagayama S;Kobayakawa K;Kobayakawa R;Tanifuji M;Sakano H;Chen WR;Mori K
通讯作者：
Mori K
影响因子：
3.7
作者：
Howard, Douglas B.;Powers, Kathleen;Harvey, Brandon K.
通讯作者：
Harvey, Brandon K.