Bright and stable fluorophores for multiparameter cell analysis

Whether you perform flow cytometry, flow sorting, or microscopy, our portfolio of fluorophores – covering wavelengths from 400 nm to 800 nm – enables you to do comprehensive multicolor analysis.

To help you plan your experiment properly, the table below summarizes important characteristics of all Vio Dye fluorophores that are available for Miltenyi Biotec antibodies. 

Vio Bright V423 (Em_max 420 nm)

Vio Bright V423 is Miltenyi Biotec’s brightest dye, excited at 405 nm. In addition to high brightness, this dye shows a significantly improved signal-to-noise ratio, minimal intra-laser compensation, and a superior detection of rare antigens. 
Vio Bright V423 is a synthetic dye which is 5× brighter than VioBlue Dye, making it an excellent choice for multiparameter cell analysis. 
 

Vio Bright V423 at a glance:

• Is the brightest Vio Dye for the violet laser.
• Provides a bright alternative for the confident detection of rare cells, as well as dim and uncharacterized markers.
• Shows minimal spillover to other detection channels within the violet laser, thereby minimizing compensation effort.
• Is available in an extensive portfolio conjugated to recombinantly engineered REAfinity™ Antibodies for higher reproducibility.

Excitation and emission maxima of fluorochromes related to Vio Bright V423.

VioBlue Dye (Em_max 452 nm)

Designed to maximize the potential of a flow cytometer’s violet laser, the VioBlue Dye shows superior performance compared to many other fluorophores excited at 405 nm, including significant advances in brightness, signal-to-noise ratios, and intralaser compensation requirements. In addition, VioBlue exhibits minimal photo-induced degradation, and can consequently be used for many different applications, such as fluorescence microscopy.
 

VioBlue Dye at a glance:

• Coumarin-based dye with excitation and emission wavelengths of 400 nm and 455 nm, respectively.
• Brighter alternative to Pacific Blue™, Alexa Fluor® 405, or BD Horizon™ V450.
• Multiplexing of VioBlue with other fluorochromes is easily possible, adding to the variety of marker combination for multiparameter flow cytometry.
• Combined use with the VioGreen Dye.

For detailed information including stability, spillover, and brightness data, please visit the MACS Handbook.
 

Excitation and emission maxima of fluorochromes related to VioBlue.

VioGreen Dye (Em_max 520 nm)

Designed to maximize the potential of a flow cytometer’s violet laser, the VioGreen Dye shows superior performance compared with many other fluorophores excited at 405 nm, including significant advances in brightness, signal-to-noise ratios, and intralaser compensation requirements. In addition, VioGreen exhibits minimal levels of photo-induced degradation, and can consequently be used for many different applications, such as fluorescence microscopy.

VioGreen Dye at a glance:

• Large Stokes shift fluorophore, emitting strong fluorescence at 520 nm upon excitation at 405 nm.
• Significantly increased mean fluorescence intensities and higher stain indices than Pacific Orange™, Krome Orange™, and BD Horizon™ V500.
• Non-protein fluorophore.
• Combined use with the VioBlue Dye.
• Perfectly suited for multiparameter flow cytometry using all current flow cytometers.

For detailed information including stability, spillover, and brightness data, please visit the MACS Handbook.
 

Excitation and emission maxima of fluorochromes related to VioGreen.

VioBright B515 and Vio B515 (Em_max 514 nm)

Vio Bright B515 and Vio B515 are extremely bright dyes for the B1/FITC-channel. The proprietary Vio Bright multimerization technology results in a 4-fold increase in brightness over the traditional FITC dye. With a 25% reduced spill-over into the B2/PE-channel and high photo stability, these dyes are excellent choices for immunofluorescence microscopy applications.

Vio Bright B515 at a glance:

• Brightest Vio Dye for the blue laser.
• Excellent choice for detection of surface markers via flow cytometry, in particular for low-expressed antigens.

Vio B515 at a glance:

• Brighter alternative to FITC.
• Perfectly suited for detection of intracellular markers via flow cytometry, such as cytokines and transcription factors.
• Optimal for use in fluorescence microscopy applications with higher photostability compared to Alexa Fluor 488.

For detailed information including stability, spillover, and brightness data, please visit the MACS Handbook.
 

Vio Bright FITC (Em_max 522 nm)

Vio Bright FITC is a blue laser–excited (488 nm) revolutionary dye, which allows binding of an increased number of FITC molecules per antibody, as compared to conventional FITC conjugation. With brightness similar to PE, Vio Bright FITC expands the dimensions of multicolor flow analysis. In addition, it provides a bright alternative for confident detection of rare cells, as well as dim and uncharacterized markers.

VioBright FITC at a glance:

• Brightness similar to PE for confident detection of low-expressed and rare markers.
• Excellent bright alternative to PE for flexible multicolor panel design.
• Signal detectable in standard FITC channel with peak excitation and emission wavelength of 496 nm and 522 nm, respectively.
• Works with your standard staining protocol.

For detailed information including stability, spillover, and brightness data, please visit the MACS Handbook.
 

PerCP-Vio 700 (Em_max 704 nm)

The PerCP-Vio 700 Dye is a tandem conjugate that combines the peridinin chlorophyll protein (PerCP) and the new Vio 700 Dye to emit a strong fluorescence at 655–730 nm upon blue laser excitation at 488 nm.  This dye is suited perfectly for the B3 channel of the MACSQuant Analyzer.
 

For detailed information including stability, spillover, and brightness data, please visit the MACS Handbook.
 

PE-Vio 770 (Em_max 770 nm)

The PE-Vio 770 Dye is a tandem conjugate, like PE-Cy™7, that exploits the principle of fluorescence-resonance-energy-transfer (FRET) allowing for large Stokes shifts between the absorbed energy of a fluorescence donor (PE) and the emission wavelength of a suitable acceptor (Vio 770) dye.

PE-Vio 770 Dye at a glance:

• Excitation with the blue (488 nm) or yellow (561 nm) laser, emission in the near-infrared region at 775 nm.
• The combination of Vio 770 as the acceptor dye and an optimized chemistry have furnished a tandem dye that is characterized by a high fluorescence intensity, minimal spillover to adjacent detection channels, and low non-specific binding to non-target cells to meet the complexity of multiparameter flow cytometry.
• Higher MFI and stain index values, in addition to lower compensation settings when compared to PE-Cy7

For detailed information including stability, spillover, and brightness data, please visit the MACS Handbook.
 

PE-Vio 615 (Em_max 619 nm)

PE-Vio 615 is a tandem dye with PE as the donor dye and Vio 615 as the acceptor dye. This tandem dye is optimized for efficient donor-to-acceptor dye energy transfer, high fluorescent intensity, and low spillover into the donor dye detection channel. Designed to be a superior alternative to ECD, PE-Texas Red®, PE-efluor® 610, PE-CF594 and PE/Dazzle™ 594, this Vio Dye expands the options for flexible multicolor panel design and provides a bright dye for confident detection of dim and rare markers.

PE-Vio 615 at a glance:

• Optimal excitation with blue (488 nm), green (532 nm), and yellow green (561 nm) laser lines for maximum flexibility.
• Excellent brightness for confident detection of dim, rare, and uncharacterized markers.
• Extensive portfolio of PE-Vio 615 dye conjugated to recombinantly engineered REAfinity Antibodies for higher reproducibility.

For detailed information including stability, spillover, and brightness data, please visit the MACS Handbook.
 

APC-Vio 770 (Em_max 770 nm)

The APC-Vio 770 Dye is a tandem conjugate, like APC-Cy™7 or APC-H7, that exploits the principle of fluorescence-resonance-energy transfer (FRET) allowing for large Stokes shifts between the absorbed energy of a fluorescence donor (APC) and the emission wavelength of a suitable acceptor (Vio 770) dye.

APC-Vio 770 Dye at a glance:

• Excitation with the yellow (561 nm) or red (635 nm) laser, emission in the near-infrared region at 775 nm.
• The combination of Vio 770 as the acceptor dye and an optimized chemistry have furnished a tandem dye that is characterized by a high fluorescence intensity, minimal spillover to adjacent detection channels, and low non-specific binding to non-target cells to meet the complexity of multiparameter flow cytometry.
• Higher MFI and stain index values, in addition to lower compensation settings when compared to APC-Cy7.

For detailed information including stability, spillover, and brightness data, please visit the MACS Handbook.
 

Vio Bright R667 and Vio R667 (Em_max 668 nm)

Vio Bright R667 and Vio R667 are extremely bright dyes for the R1/APC-channel. The proprietary Vio Bright multimerization technology results in a two-fold increase in brightness over the traditional APC dye. 
 

Vio Bright R667 at a glance:

• Brighter alternative to APC.
• Brightest Vio Dye for the red laser.
• Excellent choice for surface markers, in particular for low-expressed antigens.

Vio R667 at a glance:

• Small molecule alternative to APC.
• Perfectly suited for detection of intracellular markers via flow cytometry, such as cytokines and transcription factors.
• Optimal for use in fluorescence microscopy applications with higher photostability compared to Alexa Fluor 647.

For detailed information including stability, spillover, and brightness data, please visit the MACS Handbook.
 

Vio Bright R720 and Vio R720 (Em_max 720nm)

Vio Bright R720 and Vio R720 are organic dyes that can be detected in Alexa Fluor 700 channel (R2 channel of MACSQuant® Analyzer 16). The proprietary multimerization technology underlying the Vio Bright R720 dye typically allows for a more than 2-fold increase in brightness over the non-multimerized Vio R720. This makes Vio Bright R720 an excellent choice for multiparameter cell analysis.

Vio Bright R720 at a glance:

• Enhanced brightness with significant reduction in background noise.
• Excellent choice for surface markers, in particular for low-expressed antigens.

Vio R720 at a glance:

• Increased specificity by reducing background noise in conjugation to REAfinity Recombinant Antibodies.
• Perfectly suited for intracellular markers, such as cytokines and transcription factors.

• Fluorescence spectra overview (poster)
• Fluorochromes and instruments (poster)
• Fluorochrome brightness index (application note)
• MACS Antibodies – Quality assured from lot to lot (technical paper)
• Tandem Signal Enhancer (technical paper)