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
Calcein Deep Red™	Viability	633 nm	Cy5	7

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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
Alexa Fluor 488	Labeling	488 nm	FITC	5
FITC	Labeling	488 nm	FITC	7
iFluor® 488	Labeling	488 nm	FITC	7
Oregon Green	Labeling	488 nm	FITC	7
TRITC	Labeling	488 nm	TRITC	7
PE-Alexa Fluor 594	Labeling	488 nm	Texas Red	9
PE-Alexa Fluor 750	Labeling	488 nm	Cy7	9
PE-iFluor® 594	Labeling	488 nm	Texas Red	9
PE-iFluor® 750	Labeling	488 nm	Cy7	9
PE	Labeling	488 nm	TRITC	10
PE-Alexa Fluor 647	Labeling	488 nm	Cy5	7
PE-Cy5	Labeling	488 nm	Cy5.5	10
PE-Cy5.5	Labeling	488 nm	Cy5.5	7
PE-Cy7	Labeling	488 nm	PE-Cy7	10
ReadiUse™ Preactivated PE-iFluor® 647	Labeling	488 nm	Cy5	10
PE-Texas Red	Labeling	488 nm	Texas Red	10
Alexa Fluor 555	Labeling	532 nm	TRITC	5
Cy3	Labeling	532 nm	TRITC	5
Alexa Fluor 532	Labeling	532 nm	TRITC	7
iFluor® 514	Labeling	532 nm	Cy3/TRITC (590/40)	7
iFluor® 532	Labeling	532 nm	Cy3/TRITC (590/40)	7
iFluor® 546	Labeling	532 nm	Cy3/TRITC (590/40)	7
Alexa Fluor 594	Labeling	561 nm	Texas Red	6
iFluor® 610	Labeling	561 nm	Texas Red (610/40)	6
Sulforhodamine 101 sulfonyl chloride [Texas Red®]	Labeling	561 nm	Texas Red	6
iFluor® 555	Labeling	561 nm	Cy3/TRITC (590/40)	7
iFluor® 568	Labeling	561 nm	Cy3/TRITC (590/40)	7
iFluor® 594	Labeling	561 nm	Texas Red (610/40)	7
Alexa Fluor 750	Labeling	633 nm	Cy7	4
iFluor® 750	Labeling	633 nm	Cy7 (780/60)	4
iFluor® 680	Labeling	633 nm	Custom (712/25)	5
Alexa Fluor 680	Labeling	633 nm	Cy5.5	6
Alexa Fluor 700	Labeling	633 nm	Cy7	6
Cy7	Labeling	633 nm	Cy7	6
iFluor® 647	Labeling	633 nm	Cy5 (660/40)	6
iFluor® 660	Labeling	633 nm	Cy5 (660/40)	6
iFluor® 700	Labeling	633 nm	Custom (712/25)	6
Alexa Fluor 647	Labeling	633 nm	Cy5	7
Cy5	Labeling	633 nm	Cy5	7
iFluor® 633	Labeling	633 nm	Cy5 (660/40)	7
APC-Alexa Fluor 750	Labeling	633 nm	Cy7	8
APC-Cy5.5	Labeling	633 nm	Cy5.5	8
APC-Cy7	Labeling	633 nm	Cy7	5
APC-iFluor® 700	Labeling	633 nm	Cy7	8
APC-iFluor® 750	Labeling	633 nm	Cy7	8
APC	Labeling	633 nm	Cy5	9
iFluor® 790	Labeling	633 nm	Cy7 (780/60)	3
iFluor® 800	Labeling	633 nm	Custom (840/20)	3
iFluor® 810	Labeling	633 nm	Custom (840/20)	3
iFluor® 820	Labeling	633 nm	Custom (840/20)	3
iFluor® 860	Labeling	633 nm	Custom (885/40)	3

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Original created on July 19, 2019, last updated on July 19, 2019
Tagged under: iFluor dyes, AAT, APC, Alexa Fluor 488, Allophycocyanin, CytoTell, DNA, DAPI, Propidium iodide (PI), flow cytometry, flow cytometer

Location: Home / Application Notes / Relative Brightness of Fluorescent Dyes

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