Keyword	：

Keyword	：
Supplier	：Select	Product code	：
Product type	：	Discontinued product	：

Keyword	：
Supplier	：Select	Product code	：
Discontinued product	：

AntigenHelp	：	Cross reactivityHelp	：	Host animalHelp	：
ConjugationHelp	：	ApplicationHelp	：	PurificationHelp	：
AbsorptionHelp	：	IsotypeHelp	：	CloneHelp	：

Product type	：	Match condition	：	Discontinued product	：

Supplier	：Select	Product code	：
Supplier	：Select	Product code	：
Supplier	：Select	Product code	：
Supplier	：Select	Product code	：
Supplier	：Select	Product code	：
Supplier	：Select	Product code	：
Supplier	：Select	Product code	：
Supplier	：Select	Product code	：
Supplier	：Select	Product code	：
Supplier	：Select	Product code	：

CAS No	：

Supplier	：Select	Discontinued product	：

WEB page No search	：

HOME> Products> Cell Culture> Imaging dye> NucleoSeeing^™(Live Nucleus Green)

DNA-specific Cell Nucleus Live Imaging Reagent NucleoSeeing^™(Live Nucleus Green)

Date：March 12 2019Web Page No：81204

Funakoshi Co.,Ltd.

NucleoSeeing^™ is a nuclear staining reagent for live cell imaging that binds specifically to DNA and emits green fluorescence. It exhibits a high S/N values not only in animal cells and tissues, but also in Arabidopsis thaliana leaf cells, making it excellent for observing nuclear dynamics in live cells. NucleoSeeing^™ can also be applied for ☞ nucleus-specific pH sensing.

Example of use, NucleoSeeing<sup>™</sup> — Staining Examples of using NucleoSeeing^™

**☞ HeLa cell**, **☞ *Arabidopsis thaliana* leaf tissue** and **☞ Mouse brain pippocampus** were observed by live cell imaging, respectively; see each data for details.

☞ Product Information

Cell Nuclear Imaging in Living Cells
Principle
Features
Application
Example Data
Nucleus-specific pH sensing
Original Papers
Cited Papers
Video
Product Information

Cell Nuclear Imaging in Living Cells

Cell nucleus is one of the most important organelle, serving a DNA storehouse and responsible for cell division and epigenetic modulation. Therefore, monitoring nucleus-dynamics in live cells is considered a very important issue. Many cell nucleus-specific dyes have been developed. While nucleic acid-responsive blue fluorescent dyes, such as Hoechst series and DAPI have been widely used, these blue fluorescent dyes require UV excitation, which shows strong photo-toxicity for cells and not applicable to live cell imaging. Recently, several green or red fluorochromes for nucleus imaging on live cells have been commercially available, however, they have some disadvantages such as DNA/RNA specificity, or cell toxicity.

NucleoSeeing^™ is a DNA-responsive probe that has a DNA-specific binding tag and emits green fluorescence in the presence of DNA, enabling live cell imaging in cultured cells, cultured tissues, and plant cells such as Arabidopsis thaliana leaf cells. The product exhibits low cytotoxicity, high DNA-specific S/N value, and excellent cell nuclear staining. Fixed cells can also be observed, allowing for flexible planning of applications.
Furthermore, it has unique pH-dependent fluorescence properties and can be applied for nucleus-specific pH sensing. With the recent discussion on the importance of nuclear pH, it is expected to be used as a dedicated reagent for nuclear pH measurement.

※ ☞ For more information on nuclear pH measurement, please click here.

Comparison of nuclear staining reagents

Reagent Name	Optical Properties			Physical Properties			Observation Method
Reagent Name	Wavelength Ex/Em(nm)	Fluorescence	Photo-toxicity^＊	Nucleus Specificity	Cell Membrane Permeability	Cyto-toxicity	Live Cell	Fixed Cell
NucleoSeeing^™	488 / 520	Green	Low	High	Permeable	Low	Good	Good
Hoechst	350 / 461	Blue	High	High	Permeable	High	Good	Good
DAPI	350 / 461	Blue	High	High	Impermeable	Unknown	No	Good
Company X	485 / 498	Green	Low	Middle	Permeable	Unknown	Good	Good
Company Y	646 / 680	Red	Low	High	Permeable	High	Good	Good

* Photo-toxicity: cytotoxicity due to light irradiation

☞ Product Information

Go back to index

Principle

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.

Schematic diagram of NucleoSeeing^™

Go back to index

Features

Emits green fluorescence only upon DNA binding and exhibits a high S/N ratio. Since it is quenched in culture medium, it can be observed with high sensitivity even when added to medium.
※ To increase sensitivity, it is recommended to change the culture medium after staining.
Compared with conventional reagents such as Hoechst, there is almost no cytotoxicity.
When this reagent is added to culture medium, long-term imaging is possible.
The effect of FBS is small, and good staining is possible even under 10%FBS conditions.
※ In the presence of FBS, it is necessary to increase the concentration of the reagent. (Please refer ☞ Influence of FBS)
In addition to live-cell imaging, it can also be used for staining with fixed cells and observation of fixed cells after live-cell imaging, and can also be used for immunostaining.
It has been used in animal-derived cultured cells/tissues and plant cells (Arabidopsis leaf cells), and can stain nuclei with a high S/N ratio in both cases.
When observing plant cells, it is possible to visualize only nuclei without being affected by autofluorescence (Em: >615 nm) derived from chloroplasts.
Reversible staining is possible. By changing the medium after staining with this reagent, it is almost completely excreted from the cell in about 12-24 hours after changing the medium.
Excitation/Emission wavelength: 488 nm/520 nm
Since the fluorescence intensity changes depending on pH, it can be used as ☞ nuclear pH sensor by taking the fluorescence intensity ratio of 2 wavelengths (Ex 405 nm, Em 520 nm/460 nm) between pH6-8. It emits green fluorescence only upon DNA binding and shows a high S/N ratio.

☞ Product Information

Go back to index

Application

Live cell nucleus imaging of cultured cells, cultured tissues
Live cell nucleus imaging of plant cells
Cell nucleus staining of fixed cells
☞ Cell nucleus-specific pH sensing (pH 6-8)

Go back to index

Example Data

Index

☞ DNA-responsive fluorescent properties and nuclear-specific staining
☞ Cell toxicity
☞ Reversible staining
☞ Staining of various cultured cells
☞ Long-term time lapse imaging
☞ Staining of mouse brain (hippocampus) slice culture tissue
☞ Influence of FBS
☞ Staining of fixed cells
☞ Staining of Arabidopsis thaliana Guard cells
☞ Staining of Arabidopsis thaliana epidermal cells

DNA-responsive fluorescent properties and nuclear-specific staining

☞ Product Information

Cell toxicity

Cell toxicities of Hoechst33342, widely used as blue nuclear staining reagent, and NucleoSeeing^™ were tested by MTT assay. While Hoechst causes cell death at 5 μM, NucleoSeeing^™ keeps cell viability at least 5 μM.

Reversible staining

To evaluate the cellular retention, cells were treated with 1 μM Hoechst33342 and NucleoSeeing^™ for 15 min. The medium was changed to remove excess reagents, and fluorescence intensity was observed for 24 hours. While over 80% of Hoechst33342 signal was still remained, NucleoSeeing^™ signals were dramatically reduced within 12 hours. NucleoSeeing^™ can be used for reversible nucleus staining.

Various cultured cells were stained with NucleoSeeing — **Reversible staining of NucleoSeeing^™**

Description	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.
Storage	-20°C	CAS
Link	"NucleoSeeing" (Reversible Fluorescent Dye for Nucleus) in a minute!

DNA-specific Cell Nucleus Live Imaging Reagent NucleoSeeing^™(Live Nucleus Green)