EVOS Light Cube Selection Guide for EVOS and Countess II FL Instruments

DAPI (357/44 nm Excitation; 447/60 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	1,8-ANS	372	480
	4-methylumbelliferone	360	440
	7-amino-4-methylcoumarin (AMC)	351	430
	7-hydroxy-4-methylcoumarin	360	449
	ABQ	344	445
	Acridine	362	462
	Alexa Fluor 350	346	442
	Alexa Fluor 405	401	421
	AMCA	345	425
	AMCA-X	353	442
	BFP	382	448
	CellTrace Calcein Blue	373	440
	CellTrace Calcein Violet	401	451
	Calcofluor White	350	440
	Cascade Blue	377	420
	Dansyl	335	525
	DAPI	345	455
	DiFMU (pH 9.0)	357	450
	DyLight 350	353	432
	ER-Tracker Blue-White DPX	371	554
	Fast Blue	360	440
	FluoSpheres Blue	356	412
	Hoechst 33258	345	487
	Hoechst 33342	347	483
	LIVE/DEAD Fixable Blue Dead Cell Stain	344	442
	LysoSensor Blue	374	424
	LysoTracker Blue	373	422
	Marina Blue	365	460
	Methoxycoumarin	360	410
	Monochlorobimane(mBCI)	380	461
	NucBlue Fixed Cell Stain	345	455
	NucBlue Live Cell Stain	347	483
	Pacific Blue	405	455
	PBFI	360	420
	Phorwite AR	360	430
	Quinine Sulfate	349	461
	sgBFP (Super Glow BFP)	387	450
	SITS	365	460
	SpectrumBlue	405	449
	Thioflavin TCN	350	460
	Thiolyte	378	483
	True Blue	365	425
	Uvitex SFC	365	435
	Y66H	360	442

TagBFP (390/18 nm Excitation; 447/60 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	Alexa Fluor 405	401	421
	DyLight 405	400	420
	BFP	382	448
	Cascade Blue	400	420
	Pacific Blue	410	455
	SpectrumBlue	400	450
	TagBFP	402	457

CFP (445/45 nm Excitation; 510/42 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	Auramine O	445	500
	CellLight CFP	434	477
	ECFP	434	477
	Evans Blue	460	515
	Lucifer yellow	428	544

GFP (470/22 nm Excitation; 510/42 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	5-carboxyfluorescein(5-FAM)	492	518
	Acridine Orange + DNA	500	526
	ActinGreen (Alexa Fluor 488 phalloidin)	496	518
	Alexa Fluor 430	432	540
	Alexa Fluor 488	496	518
	ATTO 465	453	507
	Auramine	460	540
	Aurophosphine	470	540
	BCECF	499	540
	BODIPY 492/515	490	540
	BODIPY 505/515	502	540
	BODIPY FL	502	540
	BTC	464	540
	Calcein	494	540
	Calcium Green-1	506	540
	Catskill Green 540	482	540
	CellLight GFP	488	507
	CellEvent Caspase-3/7 Green	488	507
	CellTracker Green	490	540
	CellRox Green	485	520
	CFDA	494	540
	CFP	434	540
	Click-iT EdU Alexa Fluor 488	496	518
	Cy2	492	540
	CyQUANT Direct (CyQUANT GR)	500	540
	DAF-FM	493	540
	DiO (Vybrant DiO)	489	506
	Diversa Green-FP	484	507
	DyLight+C110 488	493	518
	Emerald Green	490	540
	Evans Blue	460	515
	FITC (Fluorescein)	492	520
	Fluo-3	506	540
	Fluo-4	494	540
	Fluoro-Emerald	494	518
	Fluoro-Jade B	480	525
	Fluor-X	494	520
	FluoSpheres Yellow-Green	501	513
	GFP (EGFP)	488	507
	H2DCFDA (H2-DCF,DCFR)	504	540
	HCS LipidTox Green	498	506
	HPTS (Solvent Green 7)	455	510
	LIVE/DEAD Fixable Green	498	526
	Lucifer Yellow	423	543
	LysoSensor Green	448	503
	LysoTracker Green	504	511
	Magnesium Green	506	530
	mHoneydew	478	561
	MitoTracker Green FM	490	512
	NBD Amine	450	530
	NBD-X	467	538
	NeuroTrace 500/525 Green	497	524
	Nitrobenzoxadiazole	465	510
	NucGreen Dead Cell Stain	504	523
	Oregon Green 488	498	524
	Oregon Green 514	512	532
	pHrodo Green	509	533
	Premo FUCCI Cell Cycle Sensor (S/G2/M phases)	474	509
	Qdot 525	UV	525
	Qdot 545	UV	543
	Quant-iT PicoGreen	502	522
	Rhodamine 110	496	520
	Rhodamine 123	507	529
Rhodamine Green	497	523
Rhodol Green	496	523
rsGFP (red shifted GFP, S65T)	498	516
S65C	479	507
S65L	484	510
S65T	488	511
sgGFP (Super Glow GFP)	474	509
Sodium Green	506	532
SpectrumGreen	498	522
Surf Green EX	469	526
SYBR Gold	495	539
SYBR Green I	497	521
SYBR Safe	507	524
SYTO 9	482	500
SYTO 11	506	525
SYTO 13	487	505
SYTO 16	490	520
SYTO 45	452	484
SYTO RNASelect	503	527
SYTOX Green	504	523
TET	520	541
TO-PRO-1	515	530
TOTO-1	513	531
Vybrant DyeCycle Green	505	532
wtGFP (wild type GFP, non-UV excitation)	475	509
YFP	512	529
YO-PRO-1	491	507
YOYO-1	491	509

YFP (500/24 nm Excitation; 524/27 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	Acridine Orange + DNA	500	526
	EYFP	514	527

RFP (531/40 nm Excitation; 593/40 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	ActinRed 555 (rhodamine phalloidin)	540	565
	Alexa Fluor 532	532	554
	Alexa Fluor 546	561	572
	Alexa Fluor 555	555	565
	Amplex Red	571	583
	Amplex UltraRed	568	583
	BOBO-3	570	602
	BODIPY TMR	544	569
	Calcium Orange	549	575
	Carboxy SNARF-1	548	587
	CellLights RFP	552	585
	CellMask Orange	556	571
	CellRox Orange	545	565
	CellTracker Orange	547	575
	CellTracker Red	585	612
	Cy3	552	570
	Cy3.5	579	597
	DiI (CellTracker DiI)	551	568
	DsRed Monomer	556	586
	DsRed2	563	582
	DsRed-Express	557	579
	dTomato	554	581
	DyLight 594	562	576
	Eosin	524	545
	Ethidium Bromide	521	602
	Ethidium Homodimer-1 (EthD-1)	530	618
	FluoSpheres Orange	539	560
	FocalCheck Double Orange	540	555
	HCS LipidTox Red	582	615
	HEX	533	558
	JOJO-1	529	545
	LIVE/DEAD Fixable Red	593	613
	mBanana	540	553
	MitoTracker Orange CMTMRos(MitoTracker Orange CM-H2TMRos)	551	576
	mOrange	548	568
	mTangerine	568	585
	Nile Red	551	636
	OFP	546	568
	pHrodo Red	559	586
	Phycoerythrin (PE, R-phycoerythrin)	567	576
	POPO-3	532	573
	PO-PRO-3	539	570
	Premo FUCCI Cell Cycle Sensor (G1 phase)	555	584
	Propidium Iodide (PI)	530	618
	Resorufin	570	585
	RFP	552	585
	Rhod-2	551	576
	Rhodamine Phalloidin	557	576
	Rhodamine Red	560	580
	ROX (6-ROX)	568	595
	SpectrumOrange	559	588
	Sulforhodamine 101	578	593
	SYTOX Orange	567	570
	TagRFP	555	584
	TAMRA	565	580
	TET	521	542
	Tetramethylrhodamine (TRITC)	555	580
	TurboRFP	553	574
	Vybrant DyeCycle Orange	518	562
	X-Rhod-1	570	576

Texas Red (585/29 nm Excitation; 624/40 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	2-dodecylresorufin	582	595
	5-ROX	577	603
	7-AAD	549	644
	Alexa Fluor 568	578	603
	Alexa Fluor 594	590	618
	Alexa Fluor 610	610	629
	Amplex Red	570	583
	Amplex Ultra Red	567	582
	BOBO-3	571	606
	BODIPY 630/650-X	625	641
	BODIPY TR-X	588	621
	Calcium Crimson	589	608
	CellTrace BODIPY TR methyl ester	597	625
	CellTrace calcein red-orange	575	592
	CellTracker Red CMTPX	584	613
	Click-iT Alexa Fluor 594	590	618
	Cy 3.5	581	597
	DsRed2	563	588
	DyLight 594	593	618
	FluoSpheres Crimson microspheres	620	646
	FluoSpheres Red microspheres	577	608
	HCS CellMask Red stain	587	614
	HCS LipidTOX Red	582	615
	HCS NuclearMask Red	624	644
	HcRed	594	649
	Katushka (Turbo FP635)	588	635
	LIVE/DEAD Fixable Red Dead Cell stain	595	613
	LysoTracker Red	573	592
	Magic Red	570	630
	MitoTracker Red CMXRos	578	597
	mCherry	587	615
	mKate	588	635
	mPlum	590	649
	mRaspberry	598	625
	Nile Red phospholipid	551	638
	pHrodo Red	559	587
	ReAsH (TC-ReAsH)	593	608
	rhodamine Red-X	572	591
	sulforhodamine 101	577	593
	SYTO 17	620	635
	SYTO 59	622	645
	SYTO 61	619	649
	SYTOX AADvanced dead cell stain	546	647
	Texas Red	592	615
	X-rhod-1	579	602
	YO-PRO-3	612	628
	YOYO-3	612	631

*See the complete list of CellLights Reagents

Cy5 (628/40 nm Excitation; 692/40 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	Alexa Fluor 633	631	650
	Alexa Fluor 635	634	647
	Alexa Fluor 647	650	670
	Alexa Fluor 660	663	691
	Alexa Fluor 680	679	704
	APC (Allophycocyanin)	651	660
	BODIPY 630/650X	626	641
	BODIPY 650/665	647	665
	Carboxynaphthofluorescein	600	674
	CellMask Deep Red plasma membrane stain	658	676
	CellRox Deep Red	640	665
	Click-iT Alexa Fluor 647	650	670
	Cy5	648	666
	Cy5.5	678	696
	DDAO	648	658
	DiD (Vybrant DiD)	645	670
	DRAQ5	650	680
	DyLight 633	638	658
	DyLight 649	654	673
	FluoSpheres crimson microspheres	621	646
	FluoSpheres dark red microspheres	657	683
	FxCycle Far Red	641	657
	HCS CellMask Deep Red stain	649	670
	HCS LipidTOX Deep Red	634	652
	HCS NuclearMask Deep Red	635	687
	HCS NuclearMask Red	622	644
	LIVE/DEAD Fixable Far Red	650	665
	LysoTracker Deep Red	647	668
	MitoTracker Deep Red FM	641	661
	NucRed Dead 647	642	661
	NucRed Live 647	638	686
	SYTO 17	619	638
	SYTO 59	621	644
	SYTO 60	649	681
	SYTO 62	650	681
	SYTOX Red	640	658
	TO-PRO-3	642	657
	TOTO-3	642	661
	Tubulin Tracker Deep Red	652	669
	Vybrant DyeCycle Ruby	638	686

Cy5.5 (655/46 nm Excitation; 794/16 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	Alexa Fluor 660	663	691
	Alexa Fluor 680	679	702
	Alexa Fluor 700	696	719
	ATTO 635	635	658
	ATTO 647	644	670
	ATTO 655	663	683
	ATTO 680	680	700
	ATTO 700	699	719
	Cy5.5	672	690
	DY-675	675	699
	DyLight 680	676	705
	FluoSpheres Dark Red fluorescent microspheres	656	683
	HiLyte Fluor 680	688	702
	IRDye 680 NHS Ester	683	710
	IRDye 700 Phosphoramidite	691	699
	Qdot 705 Nanocrystals	300	702
	Quasa 705	688	706
	SYTO 60	650	681

Cy7 (710/40 nm Excitation; 775/46 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	CellVue NIR780	743	776
	Cy7	745	766
	DyLight 750	752	778
	Indocyanine Green	768	807
	IRDye 800CW	774	789
	LIVE/DEAD Fixable near-IR	750	775
	XenoLight CF770	770	797

Note: Near Infrared dyes (e.g., IRDye 800CW) work best with low-magnification, high-NA objectives to maximize detection sensitivity.

CFP/YFP (445/45 nm Excitation; 542/27 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	CFP	434	477	Control YFP Emission

Notes: CFP-YFP FRET control. CFP excitation coupled with a YFP emission filter. For FRET sensitized-emission donor-acceptor bleedthrough measurements. Requires CFP and YFP light cubes for CFP-YFP FRET experiments.

AO (442/46 nm Excitation; 488LP nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	Acridine Orange + RNA	460	650
	Acridine Orange + DNA	500	526
	Alexa Fluor 430	434	540
	AmCyan	548	489
	Auramine O	460	550
	CTC Formazan	450	630
	Di-8 ANEPPS	468	635
	DiA (FAST DiA)	491	613
	Lucifer Yellow	428	544
	LysoSensor Green (pH 5)	442	505
	PerCP	488	675

Notes: Acridine Orange, longpass emission. To visualize Acridine Orange in green and orange/red emission wavelengths, we recommend using this light cube with the EVOS FL color or EVOS FL Auto cell imaging systems. The AO cube passes simultaneous green, yellow, and red fluorescence, which requires color camera detection. Works well with other fluorophores listed below in either monochrome or color camera EVOS FL cell imaging systems, provided the sample is labeled with only one dye.

AOred (442/46 nm Excitation; 628/32 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	Acridine Orange + RNA	460	650
	Acridine Orange + DNA	500	526	*
	CTC Formazan	450	630
	Di-8 ANEPPS	468	635
	DiA	491	613
	Fura Red  (high Ca2+)	436	637
	Lucifer yellow	428	544

Notes: Acridine Orange, red emission. Can be used with either EVOS FL color or EVOS FL monochrome Cell Imaging Systems. Detects red fluorescence, which can be visualized with either a color or a monochrome camera.

*Poor spectral match. Use the GFP or YFP light cube instead.

Qdot 525 (445/45 nm Excitation; 525/15 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	Acridine Orange (+DNA)	500	526
	Alexa Fluor 430	432	540
	Alexa Fluor 488	496	518
	BCECF (pH 5.2)	499	540
	BOBO-1	570	602
	Calcein	494	540
	Carboxyfluorescein (5-FAM)	492	518
	CellTracker Green CMFDA	490	540
	CellTracker Violet BMQC	415	516
	CFP	434	540
	Cy2	492	540
	CyQUANT Direct	500	540
	DAF-FM	493	540
	DiA	456	590
	DiO	484	501
	F2N12S	405	530, 585
	FITC	492	520
	Fluo-4	494	540
	FM 1-43	510	626
	GFP	488	507
	HCS CellMask Green	493	516
	LIVE/DEAD Fixable Green	498	506
	LIVE/DEAD Fixable Yellow	405	575
	Lucifer Yellow	423	543
	LysoSensor Green	448	503
	LysoSensor Yellow/Blue (pH 8.0)	329	440
	LysoTracker Green	504	511
	MitoTracker Green FM	490	512
	NBD-X	467	538
	NeuroTrace 500/525 Nissl stain	497	524
	Oregon Green 488	498	524
	Pacific Orange	440	551
	POPO-1	434	456
	PO-PRO-1	435	455
	Qdot 525	UV	525
	Rhodamine 110	496	520
	Rhodamine Green	497	523
	SYBR Gold	495	539
	SYBR Green I	497	521
	SYTO 9	482	500
	SYTO 13	487	505
	SYTO 16	490	520
	SYTO 45	452	484
	SYTO RNASelect	503	527
	SYTOX Blue	444	480
	ThiolTracker Violet	405	526
	YO-PRO-1	491	507
	YOYO-1	491	509

Qdot 545 (445/45 nm Excitation; 545/22 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	Alexa Fluor 430	432	540
	BCECF (pH 5.2)	499	540
	BOBO-1	570	602
	Calcein	494	540
	CellTracker Green CMFDA	490	540
	CellTracker Violet BMQC	415	516
	CFP	434	540
	Cy2	492	540
	CyQUANT Direct	500	540
	DiA	456	590
	DiO	484	501
	F2N12S	405	530, 585
	FITC	492	520
	FM 1-43	510	626
	GFP	488	507
	LIVE/DEAD Fixable Yellow	405	575
	Lucifer Yellow	423	543
	LysoSensor Green	448	503
	LysoSensor Yellow/Blue (pH 8.0)	329	440
	MitoTracker Green FM	490	512
	NBD-X	467	538
	NeuroTrace 500/525 Nissl stain	497	524
	Oregon Green 488	498	524
	Pacific Orange	440	551
	Qdot 545	UV	545
	Rhodamine 110	496	520
	Rhodamine Green	497	523
	SYBR Gold	495	539
	SYBR Green I	497	521
	SYTO 9	482	500
	SYTO 13	487	505
	SYTO 16	490	520
	SYTO 45	452	484
	SYTO RNASelect	503	527
	ThiolTracker Violet	405	526
	YO-PRO-1	491	507
	YOYO-1	491	509

Qdot 565 (445/45 nm Excitation; 565/16 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	Alexa Fluor 430	432	540
	BCECF (pH 5.2)	499	540
	CellTracker Violet BMQC	415	516
	CFP	434	540
	Cy2	492	540
	Di-8-ANEPPS	465	635
	DiA	456	590
	F2N12S	405	530, 585
	FM 1-43	510	626
	LIVE/DEAD Fixable Yellow	405	575
	Lucifer Yellow	423	543
	LysoSensor Green	448	503
	LysoSensor Yellow/Blue (pH 8.0)	329	440
	NBD-X	467	538
	NeuroTrace 500/525 Nissl stain	497	524
	Pacific Orange	440	551
	Qdot 565	UV	565
	SYBR Gold	495	539
	SYBR Green I	497	521
	SYTO 9	482	500
	SYTO 13	487	505
	SYTO 16	490	520
	SYTO 45	452	484
	SYTO RNASelect	503	527
	ThiolTracker Violet	405	526
	YOYO-1	491	509

Qdot 585 (445/45 nm Excitation; 586/20 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	Alexa Fluor  430	432	540
	BCECF (pH 5.2)	499	540
	CellTracker Violet BMQC	415	516
	Di-8-ANEPPS	465	635
	DiA	456	590
	F2N12S	405	530, 585
	FM 1-43	510	626
	Fura Red (+Ca2+)	436	655
	LIVE/DEAD Fixable Yellow	405	575
	Lucifer Yellow	423	543
	NBD-X	467	538
	Nile red (triglyceride)	552	636
	Pacific Orange	440	551
	Qdot 585	UV	585
	SYPRO Ruby	450	618
	ThiolTracker Violet	405	526

Qdot 605 (445/45 nm Excitation; 505/15 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	Alexa Fluor  430	432	540
	BCECF (pH 5.2)	499	540
	Di-8-ANEPPS	465	635
	DiA	456	590
	F2N12S	405	530, 585
	FM 1-43	510	626
	Fura Red (+Ca2+)	436	655
	Fura Red (-Ca2+)	473	670
	LIVE/DEAD Fixable Yellow	405	575
	Lucifer Yellow	423	543
	NBD-X	467	538
	Nile red (triglyceride)	552	636
	Pacific Orange	440	551
	Qdot 605	UV	605
	RH 795	530	712
	SYPRO Ruby	450	618

Qdot 625 (445/45 nm Excitation; 625/15 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	Di-8-ANEPPS	465	635
	DiA	456	590
	F2N12S	405	530, 585
	FM 1-43	510	626
	Fura Red (+Ca2+)	436	655
	Fura Red (-Ca2+)	473	670
	Lucifer Yellow	423	543
	Qdot 625	UV	625
	RH 795	530	712
	SYPRO Ruby	450	618

Qdot 655 (445/45 nm Excitation; 655/15 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	Acridine Orange (+RNA)	460	650
	Di-8-ANEPPS	465	635
	DiA	456	590
	FM 1-43	510	626
	FM 4-64	558	734
	Fura Red (+Ca2+)	436	655
	Fura Red (-Ca2+)	473	670
	Qdot 655	UV	655
	RH 795	530	712
	SYPRO Ruby	450	618

Qdot 705 (445/45 nm Excitation; 705/40 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	Di-8-ANEPPS	465	635
	FM 4-64	558	734
	Fura Red (+Ca2+)	436	655
	Fura Red (-Ca2+)	473	670
	Qdot 705	UV	705
	RH 795	530	712
	SYPRO Ruby	450	618

Qdot 800 (445/45 nm Excitation; 785/62 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	FM 4-64	558	734
	Qdot 800	UV	800

Qdot 525-800 (445/45 nm Excitation; 525-800 nm Emission)

	Dye	Abs Peak	Em Peak	Spectral Match
	Acridine Orange (+DNA)	500	526
	Acridine Orange (+RNA)	460	650
	Alexa Fluor  430	432	540
	Alexa Fluor  488	496	518
	BCECF (pH 5.2)	499	540
	BOBO-1	570	602
	Calcein	494	540
	Carboxyfluorescein (5-FAM)	492	518
	CellTracker Green CMFDA	490	540
	CellTracker Violet BMQC	415	516
	CFP	434	540
	Cy2	492	540
	CyQUANT Direct	500	540
	DAF-FM	493	540
	Di-8-ANEPPS	465	635
	DiA	456	590
	DiO	484	501
	F2N12S	405	530, 585
	FITC	492	520
	Fluo-4	494	540
	FM 1-43	510	626
	FM 4-64	558	734
	Fura Red (+Ca2+)	436	655
	Fura Red (-Ca2+)	473	670
	GFP	488	507
	HCS CellMask Green	493	516
	LIVE/DEAD Fixable Green	498	506
	LIVE/DEAD Fixable Yellow	405	575
	Lucifer Yellow	423	543
	LysoSensor Green	448	503
	LysoSensor Yellow/Blue (pH 8.0)	329	440
	LysoTracker Green	504	511
	MitoTracker Green FM	490	512
	NBD-X	467	538
	NeuroTrace 500/525 Nissl stain	497	524
	Nile red (triglyceride)	552	636
	Oregon Green 488	498	524
	Pacific Orange	440	551
	POPO-1	434	456
	PO-PRO-1	435	455
	Qdot 525	UV	525
	Qdot 545	UV	545
	Qdot 565	UV	565
	Qdot 585	UV	585
	Qdot 605	UV	605
	Qdot 625	UV	625
	Qdot 655	UV	655
	Qdot 705	UV	705
	Qdot 800	UV	800
	RH 795	530	712
	Rhodamine 110	496	520
	Rhodamine Green	497	523
	SYBR Gold	495	539
	SYBR Green I	497	521
	SYPRO Ruby	450	618
	SYTO 9	482	500
	SYTO 13	487	505
	SYTO 16	490	520
	SYTO 45	452	484
	SYTO RNASelect	503	527
	SYTOX Blue	444	480
	ThiolTracker Violet	405	526
	YO-PRO-1	491	507
YOYO-1	491	509