Reversible Fluorescent Dye for Cell Nucleus NucleoSeeing | Funakoshi

Reversible Fluorescent Dye for Nucleus
NucleoSeeing

NucleoSeeing is a novel live cell imaging probe which emits green fluorescence by binding to DNA specifically.
NucleoSeeing can be used for not only in animal cells or tissues, but also Guard cell of Arabidopsis thaliana with high S/N ratio. Moreover, NucleoSeeing can be used as a pH sensor in nucleus.

This product has been commercialized under the license from Nagoya Institute of Technology.

NucleoSeeing is composed of green fluorescent dye and DNA binding tag. Under DNA free condition, Nucleoseeing is in folded conformation and no fluorescent is emitted. However, once it binds to DNA, the conformation will change and emitts green fluorescence.

Fig.1 Staining of nucleus in various cultured cells
Six cell lines were treated with 1 μM of NucleoSeeing for 15 min and observed under live condition.

Fig.2 Reversible staining of NucleoSeeing
After HeLa cells were treated 1 μM Hoechst33342 and NucleoSeeing for 15 min, cells were washed by PBS and cultured for 24 hours.
While over 80% of blue signal of Hoechst33342 was still remained, green signals of NucleoSeeing were dramatically reduced within 12 hours.

Fig.3 Staining of Arabidopsis thaliana Guard cells
Arabidopsis thaliana leaves were treated with 20 μM NucleoSeeing in 10 mM MES-KOH, 50 mM KCl (pH 6.2) buffer for 60 min and observed under living condition (Ex. 488 nm and Em. 490-555 nm for NucleoSeeing, Em. >615 nm for chloroplast as autofluorescence).
Green signals from NucleoSeeing were clearly separated from plant autofluorescence derived from chloroplast.

1. Nakamura, A., et al., Chem. Commun., 50, 6149-6152 (2014)
Hoechst tagging: a modular strategy to design synthetic fluorescent probes for live-cell nucleus imaging.

2. Ueda, M., et al., ACS Cent. Sci., 3 (5), 462-472 (2017)
Noncanonical function of a small-molecular virulence factor coronatine against plant immunity: an in vivo raman imaging approach.

3. Nakamura, A. and Tsukiji, S., Bioorg. Med. Chem. Lett., 27 (14), 3127-3130 (2017)
Ratiometric fluorescence imaging of nuclear pH in living cells using hoechst-tagged fluorescein.

* hoeAc2FL in above 3 papers is equal to NucleoSeeing.

Please down load the movie from here.