8 new fluorescent live cell imaging tools – discover SPY-probes

With the great success of the SIR-probes for efficient labeling of the cell Cytoskeleton, DNA and Lysosome, Spirochrome have just launched a series of 8 new live cell fluorescent tools : The SPYTM probes.

Fig.1: New SPYTM probes wavelength profil 

These 8 new SPYTM probes, recently developed by Spirochrome, are new colour tools (green, orange, red and far red – fig.1) allowing efficient labelling of DNA (nucleus), Actin and Cytoskeleton on living and fixed cells. By keeping all the valued properties of the SIR-probes (non cytotoxic, cell permeable, highly selective, STED/SIM Compatible (fig. 2)), these 8 probes allow strong and accurate labeling of their respective targets without the need for genetic manipulation or over-expression of fluorescent proteins. Thanks to the new SPYTM probes, you will have new labeling possibilities to improve the quality of your data.

Fig.2: Compatibility of new SPYTM probes STED microscopy
FLIM filtered STED images were taken on a STED SP8 FALCON from Leica

Find out more details of these 8 new SPYTM probes below.

DNA labelling

SPY505-DNA

Fig.3 : SPY505-DNA labelling
Left: HeLa cell stained with 1x SPY505-DNA / Right: HeLa cell stained with 1x SPY505-DNA (green) and 1x SPY555-actin (magenta)

This bright green & non toxic live cell nuclear stain allows quick labeling of DNA in live & fixed cells with high specificity and very low background (fig. 3). Its absorbance and emission spectra is similar to fluorescein or Alexa488TM : (EX. max: 512 nm; Em. max: 531nm).

The SPY505-DNA can be imaged with standard fluorescein (FITC) or YFP filter sets and can be used for widefield, confocal, SIM or STED imaging in living or fixed cells and tissue. It is compatible with multicolour imaging when combined with the other SPYTM probes (SPY555, SPY595, SPY650, SPY700), the SiR-probes series and RFP or mcherry (fig. 3).

SPY555-DNA

Fig.4: HeLa cell stained with 1x SPY555-DNA

This is a bright & non toxic live cell nuclear stain allowing quick labeling of DNA in live & fixed cells with high specificity and very low background (fig. 4). Its absorbance and emission spectra are similar to tetramethyl-rhodamine (TMR): (e.g. max: 555 nm; Em. max: 580 nm).

SPY555-DNA can be imaged with standard TMR or Cy3 filter sets and it can be used for widefield, confocal, SIM or STED imaging in living or fixed cells and tissue. It allows multicolor imaging with any of the other SPYTM probes (SPY505, SPY595, SPY650, SPY700), the SiR-probes series and  the GFP.

SPY595-DNA:

Fig.5: SPY595-DNA labelling
Left: HeLa cells stained with 1x SPY595-DNA/ Right: Live HeLa cells stained with SPY555-actin (magenta), SPY595-DNA (orange) and SPY650-tubulin (blue)

This bright red & non toxic live cell nuclear stain allows quick labeling of DNA in live & fixed cells with high specificity and very low background (fig. 5).

It can be imaged with standard Alexa594TM or Texas Red filter (e.g. max: 599 nm; Em. max: 615 nm) sets and it can be used for widefield, confocal, SIM or STED imaging in living or fixed cells and tissue. SPY595-DNA enables multicolour imaging with the other SPYTM probes (SPY505, SPY555, SPY650, SPY700), the SiR-probes series and  the GFP (fig. 5)

SPY650-DNA

Fig.6: SPY650-DNA labelling
LeftBJ cells stained with 1x SPY650-DNA / Right: HeLa cell expressing EGFP-tubulin (green) co- stained with 1x SPY650-DNA (red)

It is a bright far-red & non toxic live cell nuclear stain based on the SPYTM dye series allowing quick labeling of DNA in live & fixed cells with high specificity and very low background (fig. 6). It’s emission in the red minimizes phototoxicity and sample autofluorescence: (e.g. max: 652 nm; Em. max: 674 nm). The major improvement of this SPY650-DNA is that at equal concentrations, it stains cell nuclei much better than SiR-DNA (10 fold brighter without increased toxicity) (fig.7).

It can be imaged with standard Cy5 filter sets and it can be used for widefield, confocal, SIM or STED imaging in living or fixed cells and tissue. SPY650-DNA enables multicolour imaging with the other SPYTM probes (SPY505, SPY555, SPY595, SPY700), the GFP or m-cherry (fig.6).

Fig. 7: HeLa cell stained with SPY650-DNA (Left) and SIR-DNA (right)

SPY700-DNA

Fig. 8: HeLa cell stained with SPY650-DNA (Left) and SIR-DNA (right)

This is a bright far-red & non toxic live cell nuclear stain, allowing a quick labeling of DNA in live & fixed cells with high specificity and very low background (fig. 8). It appears that at equal concentration it stains cell nuclei much better than SiR700-DNA (3-6 fold brighter without increased toxicity).

Its absorbance and emission are similar to Cy5.5: (e.g. max: 696 nm; Em. max: 718 nm). The SPY700-DNA can be imaged with Cy5 filter sets but using a 700nm LP emission filter is best and it can be used for widefield, confocal, SIM or STED imaging in living or fixed cells and tissue.

Tubulin labelling

SPY555-tubulin

Fig. 9: SPY555-Tubulin labelling
Left: HeLa cells stained with 1x SPY555-tubulin / Right: STED image of Live HeLa cell stained with SPY555-tubulin (magenta), SPY595-DNA (orange) and SiR-actin (white) with a single 775 nm STED line

It’s a bright & non toxic live cell microtubule, allowing a quick labeling of microtubules in live cells with high specificity and very low background (fig. 9). It’s absorbance and emission spectra is similar to tetramethyl-rhodamine (TMR): (Ex. max: 555 nm; Em. max: 580 nm).

It can be imaged with standard TMR or Cy3 filter sets and it can be used for widefield, confocal, SIM or STED imaging in living cells and tissue and the SPY555-Tubulin enables multicolor imaging with the other SPYTM probes (SPY505, SPY595, SPY650, SPY700), the SIR-probes series and the GFP (fig. 9).

SPY650-tubulin

Fig. 10: SPY650-Tubulin labelling
Left: HeLa cells stained with 1x SPY650-tubulin / Right: Live and dividing HeLa cell stained with SPY555-actin (magenta), SPY595-DNA (orange) and SPY650-tubulin (blue)

This is a bright far-red & non toxic live cell  microtubule stain, allowing quick labeling and brighter fluorescence, less background and which works at much lower concentrations than SiR-tubulin. Its absorbance and emission spectra are similar to Cy5: (e.g. max: 652 nm; Em. max: 674 nm) (fig. 10).

The SPY650-tubulin can be imaged with standard Cy5 filter sets and it can be used for widefield, confocal, SIM or STED imaging in living cells and tissue, and enables a multicolor imaging with with the other SPYTM probes (SPY505, SPY555, SPY595, SPY700), the GFP or m-cherry (fig. 10).

Actin labelling

SPY555-actin

Fig. 11: SPY555-Actin labelling
Left: HeLa cells stained with 1x SPY555-actin/ Right: HeLa cells stained with SPY555-acti (magenta), SPY595-DNA (orange) and SPY650-tubulin (white)

It is a bright & non toxic live F-actin stain, allowing a quick labeling of F-actin in live & fixed cells with high specificity and very low background (fig. 11). Its absorbance and emission spectra similar to tetramethyl-rhodamine (TMR): (e.g. max: 555 nm; Em. max: 580 nm).

The SPY555-Actin can be imaged with standard TMR or Cy3 filter sets and it can be used for widefield, confocal, SIM or STED imaging in living or fixed cells and tissue, and enables multicolor imaging with the other SPYTM probes (SPY505, SPY595, SPY650, SPY700), the SIR-probes series and the GFP (fig.11).


Looking for new labeling opportunities, without sacrificing the quality of your data?

Have also a look on all the other Spirochrome labeling solutions and our Live Cell Imaging interactive web page to find the good probe for your experiments.

Frédéric Samazan
Written by Frédéric Samazan
Frédéric Samazan is a Product Manager at tebu-bio, and an avid football fan in his spare time.