Flow cytometry protocols handbook
Protocols that fit your needs in flow cytometry ranging from sample preparation to numerous cell stimulation conditions, staining, immunophenotyping, and data analysis strategies.
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| 4 | 488 | 810/25 | 493 | 806 | (in buffer) 2 | flow cytometry |
Invitrogen NovaFluor Blue 800 dye has a unique emission spectrum when excited by the 488 nm laser, which fits in a spectral space that is currently not accessible by other spectral dyes. Additionally, its narrow excitation spectrum can be paired with that of NovaFluor Yellow 810 dye to replace the broad excitation spectrum of PE/Fire 810 dye. Adding spectrally unique dyes may allow for building out larger panels with minimal loss of resolution.
NovaFluor Blue 800 dye should generally be paired with moderately to highly expressed antigens in multicolor panels. The macromolecule-based NovaFluor Blue 800 dye produces highly stable fluorescence, and stained samples retain their fluorescence intensity and spectral signature when stored at 4°C. Use NovaFluor dyes with CellBlox Plus Blocking Buffer to reduce background and to block non-specific binding of NovaFluor labels, PE and APC tandems observed with macrophages and monocytes.
Additional products
- Narrow emission spectrum and minimal cross-laser excitation for better resolution
- Pair with NovaFluor Yellow 810 to replace PE/Fire 810’s broad excitation profile
- Should be paired with moderately expressed antigens to minimize spillover
- Produces stable fluorescence and is compatible with all buffers tested
Click image to enlarge
Spectral signature of NovaFluor Blue 800 dye. Normal human peripheral blood cells stained with anti-CD4 antibody (clone SK3) conjugated to NovaFluor Blue 800 dye were used for analysis. Data was acquired on a 5-laser Cytek Aurora system.
NovaFluor Blue 800 dye can be paired with NovaFluor Yellow 810 dye while providing resolution and low spillover spread. Human peripheral blood cells (PBMCs) were stained with CD27 NovaFluor Blue 800 dye and CD8 NovaFluor Yellow 810 dye. Viable cells were used for analysis, as determined by LIVE/DEAD Fixable Blue Dead Cell Stain Kit, for UV excitation. Data was collected on a 5-laser Cytek Aurora Flow Cytometer.
NovaFluor dyes
NovaFluor dyes are built using Phiton technology and are compatible with both spectral flow cytometry and traditional flow cytometry. NovaFluor dyes exhibit narrow excitation spectra and minimal cross-laser excitation, which helps reduce spectral spillover for better marker resolution. In addition, their unique spectral signatures can provide the opportunity to detect additional markers in flow cytometry panels by opening up previously unusable channels.
Additional resources
Fluorophore and reagent selection guide for flow cytometry
A handy reference poster featuring the broad range of our available dyes and labeling reagents.
Imaging protocol handbook
Protocols that fit your needs in imaging ranging from sample and assay preparation to staining, labeling, and data analysis strategies.
Fluorophore and reagent selection guide for cell imaging
A tool for selecting the optimal fluorescent dyes for your Invitrogen EVOS cell imaging systems.
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