EVOS Light Cube Selection Guide | Thermo Fisher Scientific


DAPI	RFP	CFP-YFP	Qdot 605
TagBFP	Texas Red	AO	Qdot 625
CFP	Cy5	AOred	Qdot 655
GFP	Cy7	Qdot 525	Qdot 705
YFP		Qdot 545	Qdot 800
		Qdot 565	Qdot 525-800
		Qdot 585

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
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

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

Light Cubes

Specialty Light Cubes