| RRC ID |
74561
|
| Author |
Yamauchi K, Okamoto S, Ishida Y, Konno K, Hoshino K, Furuta T, Takahashi M, Koike M, Isa K, Watanabe M, Isa T, Hioki H.
|
| Title |
Fluorochromized tyramide-glucose oxidase as a multiplex fluorescent tyramide signal amplification system for histochemical analysis.
|
| Journal |
Sci Rep
|
| Abstract |
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.
|
| Volume |
12(1)
|
| Pages |
14807
|
| Published |
2022-9-12
|
| DOI |
10.1038/s41598-022-19085-9
|
| PII |
10.1038/s41598-022-19085-9
|
| PMID |
36097273
|
| PMC |
PMC9468149
|
| MeSH |
Animals
Coloring Agents*
Fluorescent Antibody Technique
Glucose Oxidase*
In Situ Hybridization, Fluorescence / methods
Mice
Peroxidases
|
| IF |
3.998
|
| Resource |
| Human and Animal Cells |
293T(RCB2202) |
