Flow Cytometry

Flow Cytometry is a powerful technique to measure multiple parameters from cell populations. Flow cytometers differentiate cells based on their size, internal complexity, and fluorescence signature. Qualitative and quantitative analyses of cell populations have relevance for clinical and research applications.

Below is a table listing our recommended fluorophores/tandems for flow cytometry (click a dye name to view more information and/or place an order):

Excitation laser line [nm]	Fluorochromes Used^†				Superior Seta Replacements (Free dye / Ab conjugate^‡ in phosphate buffer pH 7.4)
Excitation laser line [nm]	Fluorochrome (Free dye)	λ_max Ab [nm]	λ_max Fl [nm]	Brightness at the excitation laser line [M^–1cm^–1]	Free dye / Ab conjugate	λ_max Ab [nm]	λ_max Fl [nm]	Brightness at the excitation laser line [M^–1cm^–1]	Advantage and specific feature
380	Hoechst 34580	380	438	n.a.^§	Seta-375 / IgG (D/P = 5)	372/372	476/480	20,500/50,000	Larger Stokes shift
					SeTau-380 / BSA	381/383	480/468	900/3,000	Larger Stokes shift
					Seta-385 / IgG (D/P = 4)	384/387	508/505	19,400/34,700	Larger Stokes shift
405	Pacific Blue	401	452	21,000	Seta-400 / IgG (D/P = 4)	399/401	576/569	8,000/15,100	Larger Stokes shift
	Pacific Blue	401	452	21,000	Seta-405 / IgG (D/P = 3)	401/404	452/453	20,800/49,900	Pacific Blue analog
	Pacific Orange	400	551	n.a.^§	Seta-405 / IgG (D/P = 3)	401/404	452/453	20,800/49,900	Shorter Stokes shift
	AmCyan	468	498	1,900	SeTau-405 / IgG (D/P = 2)	405/405	518/518	11,000/6,900	~6× Brighter, Larger Stokes shift
436	CCVJ	440	505	n.a.^§	Seta-470 / Glycerol:Water 10 cP, +65 °C / 700 cP, -6 °C	460/460/460	521/525/525	560/n.d.^/n.d.^	Viscosity sensitive
	Pacific Orange	400	551	n.a.^§	SeTau-425 / BSA (D/P = 2)	425/427	545 /532	1,500/1,000	Shorter Stokes shift
	AmCyan	468	498	5,100	Seta-PerCP-680	481	676	79,000	~15× Brighter. Larger Stokes shift
488	FITC	488	491	67,000	SeTau-488 / IgG (D/P = 2)	486/484	532/530	22,800/71,600	Larger Stokes shift. Highly stable
	Alexa 488	496	519	65,000	SeTau-488 / IgG (D/P = 2)	486/484	532/530	22,800/71,600	Larger Stokes shift. Highly stable
	Cy3	555	569	3,400	SeTau-500 / IgG (D/P = 2)	502/500	568/566	31,200/53,100	~9× and ~14× brighter than Cy3 and Alexa 555. Larger Stokes shift. More photo- and oxidatively stable
	Alexa 555	553	568	2,200	SeTau-500 / IgG (D/P = 2)	502/500	568/566	31,200/53,100
	BDP FL	502	511	n.a.^§	Seta-470 / Glycerol:Water 10 cP, +65 °C / 700 cP	460/460/460	521/525/525	560/n.d.^/n.d.^	Viscosity sensitive
	R-PE	565	573	988,000	R-PE	565	573	988,000	Identical
	R-PE – Cy5	565	666	285,800	Seta-R-PE-670	565	671	340,000	20% Brighter
	R-PE – Cy5.5	565	671	234,400	Seta-R-PE-670	565	671	340,000	45% Brighter
	R-PE – Cy7	565	778	148,000	Seta-R-PE-775	565	774	148,000	R-PE – Cy7 analog
	PerCP	477	678	162,400	PerCP	477	678	162,400	Identical
	PerCP – Cy5.5	488	678	80,500	Seta-PerCP-680	481	676	88,800	PerCP – Cy5.5 analog
532	Alexa 555	553	568	7,800	Seta-555 / IgG (D/P = 2)	555/556	570/571	3,400/n.d.^*
	Alexa 555	553	568	7,800	Seta-580 / BSA (D/P = 1)	578/585	602/609	15,900/4,300	~2× Brighter
	Cy3	555	569	11,700					~40% Brighter
	5(6)-ROX	578	602	15,900					5(6)-ROX analog
	mCherry	587	610	5,700					~3× Brighter
	R-PE	565	573	1,317,000	R-PE	565	573	1,317,000	Identical
	R-PE – Cy5	565	666	412,800	Seta-R-PE-670	565	671	435,900	R-PE – Cy5 analog
	R-PE – Cy5.5	565	671	324,600	Seta-R-PE-670	565	671	435,900	~30% Brighter
	R-PE – Cy7	565	778	190,500	Seta-R-PE-775	565	774	190,500	R-PE – Cy7 analog
	PerCP – Cy5.5	488	679	76,500	Seta-PerCP-680	481	676	57,000
594	5(6)-ROX	578	602	21,900	Seta-580 / BSA (D/P = 1)	578/585	602/599	21,900/6,000	5(6)-ROX analog
	mCherry	587	610	13,600	Seta-640 / IgG (D/P = 2)	636/648	651/653	4,600/19,900	Ab conjugate is ~50% brighter than mCherry
	cl-APC	652	660	263,100	cl-APC	652	660	263,100	Identical
635	Cy5	647	665	50,300	Zy5 / IgG (D/P = 2)	647/647	665/665	50,300/70,800	Identical
	Alexa 647	650	671	60,600	SeTau-647 / IgG (D/P = 2)	649/648	695/694	74,000/128,600	Larger Stokes shift, ~20% Brighter
	Alexa 700	702	723	23,100	Seta-650 / IgG (D/P = 2)	651/654	671/673	35,100/80,200	~50% Brighter
	Alexa 700	702	723	23,100	Seta-660 / IgG (D/P = 2)	661/665	672/675	4,900/38,900
	cl-APC	650	660	367,000	cl-APC	650	660	367,000	Identical
	cl-APC – Cy7	650	779	68,900	Seta-APC-780	650	780	116,700	~70% Brighter
	cl-APC – Alexa 700	650	719	132,500	Seta-APC-705	655	681	384,000	~3× Brighter
650	Cy5	650	665	67,500	SeTau-647 / IgG (D/P = 2)	649/648	695/694	122,000/210,700	~80% and ~40% brighter than Cy5 and Alexa 647, respectively, and 3-4 orders more stable towards 2% H₂O₂. Larger Stokes shift. ~12× More photostable.
	Alexa 647	650	671	89,100	SeTau-647 / IgG (D/P = 2)	649/648	695/694	122,000/210,700
	Alexa 647	650	671	89,100	SeTau-660 / IgG (D/P = 2)	663/663	694/694	94,400/162,400	~80% Brighter when conjugated to Ab. More stable to light and oxidizers.
	Cy5.5	676	694	23,500	Seta-650 / IgG (D/P = 2)	651/654	671/673	56,000/128,400	~2× Brighter.
					Seta-660 / IgG (D/P = 2)	661/665	672/675	10,700/83,800	~4× Brighter when conjugated to Ab.
					SeTau-665 / IgG (D/P = 2)	664/662	712/716	47,900/54,200	~2× Brighter. More stable to light and oxidizers
					Seta-670 / IgG (D/P = 1)	667/673	688/693	7,300/12,500
					SeTau-670 / IgG (D/P = 4)	673/672	694/693	39,600/61,600	~70% Brighter. More stable to light and oxidizers
	CypHer5E	643	660	7,000	Square-650-pH at pH 5.6 / pH 9.0	653/535	671/663	20,900/260 (3,600 at λ* = 532 nm)	pH sensitive. ~3× Brighter. Suitable for ratiometry
	cl-APC	650	660	496,000	cl-APC	650	660	496,000	Identical
	cl-APC – Alexa 700	650	719	175,000	Seta-APC-705	655	681	574,000	~3× Brighter. Larger Stokes shift. More photo- and oxidatively stable
	cl-APC–Cy7	651	779	91,000	Seta-APC-780	651	756	208,000	~2× Brighter
680	Cy5.5	676	694	50,400	Zy5.5 / IgG (D/P = 2)	676/683	694/698	50,400/43,800	Identical
					SeTau-670 / IgG (D/P = 4)	673/672	694/693	97,300/110,400	~2× Brighter. More stable to light and oxidizers
					Seta-680 / IgG (D/P = 2)	678/680	698/701	47,500/103,000	Ab conjugate is brighter than for Cy5.5
	Alexa 680	681	704	65,900	SeTau-680 / IgG (D/P = 2)	683/682	705/703	122,300/244,600	~2× Brighter. More stable to light and oxidizers
	cl-APC – Alexa 700	650	719	126,000	Seta-APC-705	655	681	651,500	~5× Brighter
700	Cy7	756	779	8,400	Zy7	749	774	8,400	Identical
	Alexa 700	696	719	50,200	Seta-700 / IgG (D/P = 2)	687/705	703/715	7,200/7,200	Seta version
	Alexa 750	752	776	16,700	Seta-750 / IgG (D/P = 2)	753/756	780/783	9,600/20,500	Seta version
750	Cy7	756	779	26,000	Zy7	749	774	26,000	Identical
750	Alexa 750	752	776	34,800	Seta-750 / IgG (D/P = 2)	753/756	780/783	31,600/68,700	Ab conjugate is ~2× brighter than for Alexa 750
780	Cy7	756	779	7,400	Zy7	749	774	7,400	Identical
	Alexa 790	782	805	n.a.^§	Seta-780	777	807	2,600	Seta version
	APC–Cy7	651	779	70,300	Seta-APC-780	651	756	195,700	~3× Brighter

^† Fluorochromes Used: data listed for free dyes.
^‡ Fluorochrome conjugated to antibody (protein). D/P is dye-to-protein ratio.
^* Not determined.
^§ Not available — brightness, extinction coefficient, and/or fluorescence quantum yield have not been reported in the literature.

			Excitation Light Sources												Characteristics
Product Number (Specs Sheet)	Product Name (Product Info)	Target Group	380	405	436	488	532	594	635	650	680	700	750	780	Medium	λ abs [nm]	ε [M ^–1 cm^–1]	λ em [nm]	QY [%]	FLT [ns]	Buy

K7-547 K7-547	SeTau-405-NHS SeTau-405-NHS	NH₂	•	•											PB 7.4	405	13,800	518	80	9.3
K7-545 K7-545	SeTau-425-NHS SeTau-425-NHS	NH₂	•	•	•										PB 7.4	425	4,200	545	39	26.2
K8-1384 K8-1384	Seta-700-NHS Seta-700-NHS	NH₂	•	•						•	•	•			PB 7.4	687	177,000	703	6
K8-1682 K8-1682	Seta-660-di-NHS Seta-660-di-NHS	NH₂							•	•					PB 7.4	661	220,000	672	11	0.9
K8-5035 K8-5035	Seta-650-NHS Seta-650-NHS	NH₂							•	•					PB 7.4	651	200,000	671	28
K8-5036 K8-5036	Seta-650-Maleimide Seta-650-Maleimide	SH							•	•					PB 7.4	652	200,000	672	28
K8-7055 K8-7055	Seta-780-NHS Seta-780-NHS	NH₂										•	•	•	PB 7.4	777	215,000	807	1.2
K9-4119 K9-4119	SeTau-665-NHS SeTau-665-NHS	NH₂								•	•				PB 7.4	664	160,000	712	53	3.1
K9-4149 K9-4149	SeTau-647-NHS SeTau-647-NHS	NH₂							•	•					PB 7.4	649	200,000	695	61	3.2
K8-1341 K8-1341	Seta-670-Maleimide Seta-670-Maleimide	SH	•	•					•	•					PB 7.4	667	180,000	688	7
K4-205 K4-205	Seta-375-NHS Seta-375-NHS	NH₂	•												PB 7.4	372	24,500	476	86	3.1
K7-548 K7-548	SeTau-405-Maleimide SeTau-405-Maleimide	SH	•	•											PB 7.4	405	13,800	518	51	9.1
cl-APC cl-APC	Crosslinked APC Crosslinked APC							•	•	•					PB 7.4	650	700,000	662
R-PE R-PE	Lyophilized R-PE Lyophilized R-PE					•	•								PB 7.4	565	1,960,000	573
K10-6801 K10-6801	Seta-PerCP-680 Seta-PerCP-680				•	•	•								PB 7.4	481	380,000	676
K10-7801 K10-7801	Seta-APC-780 Seta-APC-780								•	•					PB 7.4	651	1,100,000	780
K10-6701 K10-6701	Seta-R-PE-670 Seta-R-PE-670					•	•								PB 7.4	488	1,100,000	671
K10-0775 K10-0775	Seta-R-PE-775 Seta-R-PE-775					•	•								PB 7.4	488	1,100,000	774
K10-7051 K10-7051	Seta-APC-705 Seta-APC-705								•	•					PB 7.4	655	1,100,000	703
K7-544 K7-544	SeTau-425-Maleimide SeTau-425-Maleimide	SH	•	•	•										PB 7.4	425	4,200	545	39	26.2
K4-225 K4-225	Seta-385-NHS Seta-385-NHS	NH₂	•												PB 7.4	384	25,000	512	78
K6-5052 K6-5052	Seta-405 NHS Seta-405 NHS	NH₂	•	•											PB 7.4	401	30,000	452	70
K4-215 K4-215	Seta-400-NHS Seta-400-NHS	NH₂	•	•											PB 7.4	399	24,000	576	34
K9-3152 K9-3152	SeTau-488-NHS SeTau-488-NHS	NH₂				•									PBS 7.4	486	59,000	532	27	1.2
K9-3153 K9-3153 ^new	SeTau-488-Maleimide SeTau-488-Maleimide	SH				•									PBS 7.4	484	59,000	531	20
K9-4159 K9-4159 ^new	SeTau-660-NHS SeTau-660-NHS	NH₂							•	•					PB 7.4	663	240,000	694	50	3.36
K9-3162 K9-3162 ^new	SeTau-500-NHS SeTau-500-NHS	NH₂				•									PB 7.4	502	59,600	567	43	2.8

Seta-405 is identical to Pacific Blue with excitation and emission at 401/452 nm.

SeTau-405 is a spectral analog of Pacific Orange with excitation and emission at 405/518 nm.

SeTau-488 with a similar absorption and emission as FITC and Alexa 488 (Φ = 0.92, ε = 73,000 M^-1cm^-1 and Φ·ε = 67,160 M^-1cm^-1) exhibits a lower brightness as a free dye, but its brightness gets increased when it is attached to a biomolecule, and it becomes an almost unquenchable fluorophore when labeled to an antibody (see specs sheet). Exc./Em. max of IgG-conjugates: 484/530 nm.

Quantum yield of SeTau-488 — IgG vs Alexa 488 — IgG

Quantum yield of SeTau-488 — IgG (K9-3152) and Alexa 488 — IgG vs. dye-to-protein ratio

Total brightness (QY × ε × D/P) of SeTau-488 — IgG (K9-3152) and Alexa 488 — IgG vs. dye-to-protein ratio

SeTau-647 is an extremely bright label unsurpassed by any other red dye for excitation with the 633 nm laser. Its Q.Y. in aqueous solutions is 60%, and its Stokes' shift is >40 nm. It is currently one of the brightest fluorescent dyes for the red channel.

Seta-APC-705 and Seta-R-PE-775 are our latest additions to this product line.

Seta-APC-780 is a significantly brighter and much more photostable replacement for APC-Cy7.

Peripheral blood samples stained with CD81 — Seta-APC-780

Peripheral blood samples stained with CD45 — Seta-APC-780

Peripheral blood samples stained with CD3 — Seta-APC-780

Plasma cells stained with CD38 — Seta-APC-780

Please contact us if you are interested in a sample for testing.