Reversible Fluorescent Dye for Cell Nucleus

news April 22 2019

NucleoSeeing is a novel live cell imaging probe which emits green fluorescence when binding specifically to DNA. NucleoSeeing can be used in animal cells or tissues, but also in e.g. Guard cell of Arabidopsis thaliana. Moreover, NucleoSeeing can be used as a pH sensor in nucleus. 

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

181-FDV-0029

Working principle

NucleoSeeing is composed of green fluorescent dye and a DNA binding tag. Under DNA free conditions, the probe is in folded conformation and not fluorescent. Once it binds to DNA, the conformation will change and a green fluorescence signal will be emitted.

Comparison table of dyes for nucleus

Ex/Em Fluorescence Photo-toxicity Nucleus Specificity Cell Mem. Permeability Cyto-toxicity Live Fixed
NucleoSeeing 488/520 Green Low Yes Yes Low Yes Yes
Hoechst 350/461 Blue High Yes Yes High Yes Yes
DAPI 350/461 Blue High Yes No N/A No Yes
Company X 485/498 Green Low No Yes N/A Yes Yes
Company Y 646/680 Red Low Yes Yes High Yes Yes

 

Features/Benefits

・ Excitation / Emission : 488 nm / 520 nm

・ High S/N ratio

・ Low cytotoxicity

・ Can be used for both live and fixed cells

・ Can be used for animal samples and plant cells.

(validated in leaf and guard cell of A. thaliana)

・ Reversible : Can be washed out by replacing the medium

・ Can be used as nucleus pH sensor

 

Example Data

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.

References

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.

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.

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.