Relative Brightness of Fluorescent Dyes
DNA Dyes
Fluorescence images of oleic acid treated HeLa cells stained with Cell Navigator® Lipid Droplets Fluorescence Assay Kit (Cat#22735) and Nuclear Green™ LCS1 (Cat#17540).
Table 1. Relative Brightness of DNA Dyes
Dye Name ▲ ▼ | Function ▲ ▼ | Laser ▲ ▼ | Filter ▲ ▼ | Brightness ▲ ▼ |
DAPI | DNA | 355 nm | DAPI | 6 |
Nuclear Blue™ DCS1 | DNA | 355 nm | DAPI | 6 |
Propidium iodide | DNA | 488 nm | Texas Red | 2 |
Nuclear Green™ | DNA | 488 nm | FITC | 6 |
Propidium iodide | DNA | 532 nm | Texas Red | 3 |
Nuclear Red™ | DNA | 633 nm | Cy5.5 | 6 |
Proliferation Dyes
Cell proliferation assay with CytoTell™ Green and CFSE. Jurkat cells (~2×106 cells/mL) were stained with CytoTell™ Green or CFSE on Day 0. The cells were passed serially at 1:1 ratio on the day specified. Fluorescence intensity was measured with ACEA NovoCyte 3000 flow cytometer in FITC channel on the day of passage. Successive generations were represented by different colors.
Table 2. Relative Brightness of Proliferation Dyes
Dye Name ▲ ▼ | Function ▲ ▼ | Laser ▲ ▼ | Filter ▲ ▼ | Brightness ▲ ▼ |
CytoTell™ Blue | Proliferation | 405 nm | DAPI | 6 |
CFSE | Proliferation | 488 nm | FITC | 7 |
CytoTell™ Green | Proliferation | 488 nm | FITC | 7 |
CytoTell™ Orange | Proliferation | 488 nm | Cy3/TRITC | 7 |
CytoTell™ UltraGreen | Proliferation | 488 nm | FITC | 7 |
CytoTell™ Red 590 | Proliferation | 561 nm | Cy3 | 7 |
CytoTell™ Red 650 | Proliferation | 633 nm | Cy5 | 7 |
Viability Dyes
Images of Live HeLa cells stained with CytoCalcein Violet 500 (Cat.22013). Cell nuclei were stained with Hoechst 33342 (Blue, Cat#17535).
Table 3. Relative Brightness of Viability Dyes
Dye Name ▲ ▼ | Function ▲ ▼ | Laser ▲ ▼ | Filter ▲ ▼ | Brightness ▲ ▼ |
Calcein Blue | Viability | 405 nm | DAPI | 6 |
Calcein UltraBlue™ AM | Viability | 405 nm | DAPI | 6 |
CytoCalcein™ Violet 450 | Viability | 405 nm | DAPI | 7 |
CytoCalcein™ Violet 500 | Viability | 405 nm | FITC | 7 |
7-AAD | Viability | 488 nm | Cy5 | 1 |
Calcein *UltraPure Grade* | Viability | 488 nm | Cy5 | 1 |
Calcein Orange™ | Viability | 488 nm | TRITC | 7 |
Calcein UltraGreen™ AM | Viability | 488 nm | FITC | 7 |
7-AAD | Viability | 532 nm | Cy5 | 2 |
Calcein Red™ AM | Viability | 561 nm | Cy3/TRITC | 7 |
Labeling Dyes
HeLa cells were incubated with (Tubulin+) or without (Tubulin-) mouse anti-tubulin followed by iFluor® 488 goat anti-mouse IgG conjugate (Green, Left) or Alexa Fluor® 488 goat anti-mouse IgG conjugate (Green, Right), respectively. Cell nuclei were stained with Hoechst 33342 (Blue).
Table 4. Relative Brightness of Labeling Dyes
Dye Name ▲ ▼ | Function ▲ ▼ | Laser ▲ ▼ | Filter ▲ ▼ | Brightness ▲ ▼ |
Alexa Fluor 350 | Labeling | 355 nm | DAPI | 2 |
iFluor® 350 | Labeling | 355 nm | DAPI | 6 |
iFluor® 450 | Labeling | 405 nm | FITC | 5 |
Pacific Green | Labeling | 405 nm | FITC | 5 |
Pacific Orange | Labeling | 405 nm | TRITC | 2 |
iFluor® 405 | Labeling | 405 nm | DAPI | 6 |
iFluor® 430 | Labeling | 405 nm | FITC | 6 |
PacBlue | Labeling | 405 nm | DAPI | 2 |
Alexa Fluor 555 | Labeling | 488 nm | TRITC | 4 |
Cy3 | Labeling | 488 nm | TRITC | 4 |
Original created on July 19, 2019, last updated on October 20, 2022
Tagged under: iFluor dyes, AAT, APC, Alexa Fluor 488, Allophycocyanin, CytoTell, DNA, DAPI, Propidium iodide (PI), flow cytometry, flow cytometer