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.
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 |
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.
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 |
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.
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.