期刊
SCIENTIFIC REPORTS
卷 12, 期 1, 页码 -出版社
NATURE PORTFOLIO
DOI: 10.1038/s41598-022-19085-9
关键词
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资金
- JSPS KAKENHI [JP20K07231, JP21H03529, JP20K07743, JP21H02589, JP20H05628, JP21H02592]
- Japan Agency for Medical Research and Development (AMED) [JP21dm0207112, JP19dm0207093, JP18dm0207020]
- Moonshot R&D from the Japan Science and Technology Agency (JST) [JPMJMS2024]
- Fusion Oriented Research for disruptive Science and Technology (FOREST) from JST [JPMJFR204D]
- Research Institute for Diseases of Old Age at the Juntendo University School of Medicine [X2016, X2001]
- Private School Branding Project
Tyramide signal amplification (TSA) is a highly sensitive method for histochemical analysis. In this study, a fluorochromized tyramide-glucose oxidase (FT-GO) was developed as a multiplex fluorescent TSA system. FT-GO enhanced immunofluorescence signals while maintaining low background signals. It showed a more widespread distribution of monoaminergic projection systems and was successfully used in multiplex fluorescent in situ hybridization.
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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