Vio Bright R720 Vio R720 fluorophores | Vio Dye fluorophores | MACS Antibodies | Products | Miltenyi Biotec

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.

Fluorophores at a glance

Overview of fluorophore characteristics

Laser	640nm
Excitation max	695 nm
Emission max	720 nm
Application	Flow cytometry
Common filter set (FC)	725/40 nm
Stain index (Vio Bright R720)	4 out of 5
Photostablility	Photostable to ambient light
Fixation stability	PFA

Laser and filter compatibility

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Absorption and emission spectra of Vio Bright R720 and Vio R720

With a maximum excitation and emission at 695 nm and 720 nm, respectively, Vio R720 and Vio Bright R720 conjugates are fully compatible with standard filter sets from all major flow cytometry hardware providers, giving researchers the flexibility to use these dyes with all existing platforms. The absorption and emission spectra of these two dyes are similar to the spectra of Alexa Fluor 700 and APC-R700.

Enhanced brightness

Brighter and clearer staining for the red laser

The multimerization technology for Vio Bright dyes results in a 2-fold or higher increase in brightness of Vio Bright R720 compared to a conventional fluorophore like Alexa Fluor 700. Increased brightness of Vio Bright R720 conjugates is observed across different antigens; the data below show examples of CD19 and CD56 staining. Greater brightness of Vio Bright R720 compared to Alexa Fluor 700 is also reflected in the values of mean fluorescence intensity (MFI) and stain index (SI) for the detection of specific cells.

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Comparison of MFI and SI for CD56 antibodies conjugated to Vio Bright R720 or Alexa Fluor 700. The table also displays the percentage of positive cells.

	Vio Bright R720 (NK)	Alexa Fluor 700 (NK)
MFI	11.37	2.68
SI	31.25	6.74
% - #	30.96	28.68

Low background

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Decreased background noise

Starting material like lysed whole blood may result in unspecific background noise, therefore it is important to avoid additional unspecific signal from reagents. Vio Bright R720 conjugates demonstrate decreased background noise compared to Alexa Fluor 700 and APC-R700.

Fixation stability

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Excellent performance with different staining protocols

Both Vio R720 and Vio Bright R720 show excellent stability after fixation with paraformaldehyde, and are compatible with different surface and intracellular staining protocols and buffer sets. Conjugate performance may vary depending on antigen nature upon fixation.

Photostability

The stability of both Vio R720 and Vio Bright R720 upon exposure to ambient light was analyzed at different time points. Analysis of the photostability of a CD4 antibody conjugated to Vio R720 and Vio Bright R720 showed no major change in both MFI and SI after up to 24 hours of continuous exposure to ambient light (~850 lux), which is similar to other commercially available alternatives.

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Resources for Vio Bright R720 and Vio R720 fluorophores

Multicolor flow cytometry panels

Miltenyi Biotec–tested panels (MBTPs) are a collection of multicolor antibody panels that are optimized and pre-designed for specific flow cytometry applications. They are developed and validated by Miltenyi Biotec’s R&D experts, as well as by the customers who actually use Miltenyi Biotec antibodies.