Cytek AuroraFluorochrome Selection Guidelines

Last Updated: October, 2018

Fluorochrome SignaturesDyes can be used in combination as long as they have a distinct spectrum signature.

In designing a multicolor panel, you should look for dyes with unique spectra and consider their impacts on other dyes (see slide 20)

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Violet Laser Excitable Dyes

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Violet Laser Excitable Dyes with Similar or Identical SignatureseFluor450 –Pacific Blue –vioBlue – Live/Dead Violet

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Super Bright 436, Zombie Violet

Violet Laser Excitable Dyes with Similar or Identical SignaturesBV510 – VioGreen– Zombie Aqua –Live/Dead Yellow – eFluor 506

e506 is slightly different

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Brilliant Violet vs Super Bright (SB): almost identical

BV421vs

SB 436

BV605vs

SB 600

BV650vs

SB 645

BV711vs

SB 702

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Qdot655

Violet Laser Excitable Dyes Unique SignaturesBV421

SB436, Zombie Violet

SB 600-BV605

Qdot605

SB 645-BV650

Qdot705

SB702, BV711

BV750

BV785

Qdot 800

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Live/Dead Blue

Pacific Blue, eFluor450, vioBlue, Live/Dead Violet

Zombie Yellow

BV570

Live/Dead Red

BV480

efluor506

BV510, VioGreen, Zombie Aqua, Live/Dead Yellow

Zombie Red

Blue Laser Excitable Dyes

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Blue Laser Excitable Dyes with Identical Signatures (1)BB515- sVio515- Vio515 AF 488, FITC, vioBright FITC, Zombie Green

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PE Dazzle594, PE eFluor 610, PE Texas Red PerCP vio700, PerCP eFluor 710

PE Vio770, PE Cy7

Blue Laser Excitable Dyes with Identical Signatures (2)

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Blue Laser Excitable Dyes Unique Signatures

BB515- sVio515- Vio515

AF 488, FITC, vioBright FITC, Zombie Green

PE

Alexa Fluor 532

PE Cy5

PE Cy55

PerCP

BB700

PerCP vio700, PerCP eFluor 710

PE Vio770, PE Cy7

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Live/Dead Green

PE Dazzle594, PE CF594, PE eFluor 610, PE Tx Red

PerCP Cy55

Red Laser Excitable Dyes

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Red Laser Excitable Dyes with Identical SignaturesAlexa 647, Vio 667, Live/Dead Far Red

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APC-Cy7, APC-H7, APC Fire 750 , APC Vio770, Live/Dead Near-IR

Red Laser Excitable Dyes with Unique Signatures

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APC

Alexa Fluor 647, Vio667, Live/Dead Far Red

APC Cy55

Alexa Fluor 700

APC H7, APC Fire750, APC Cy7, APCvio770

Zombie NIR

Example of 24 dyes that can be used in combination(CAREFUL PANEL DESIGN IS NEEDED)

Fluorophore Fluorophore Fluorophore

BB515 APC BV421

Alexa Fluor 488 or FITC Alexa Fluor 647 Super Bright 436

Alexa Fluor 532 APC-R700 or AF700 eFluor450 or equivalent

PEAPC-Fire 750 or

equivalentBV480

PE/Dazzle 594 or equivalent

BV510

PE-Cy5 BV570

PerCPCy55 BV605

PerCP-eFluor710 BV650

PE-Cy7 BV711

BV750

BV785

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Stain IndexesData generated using CD4 staining in human whole blood

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Stain Index 53 dyes

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Stain Index Violet Excitable dyes

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Stain Index Blue Excitable dyes

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Stain Index Red Excitable dyes

Spread MatrixDyes used in combination need to have a unique spectrum AND also need to be assessed in terms of spread that they introduce to other dyes.

For example PerCP Cy5.5 and PE Cy5.5 have distinct signatures, but because of emission in the same wavelength range and significant spread introduced by PE Cy5.5, you will use one or the other.

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To read this table: fluor in the row impacts the one in the column. Red means the fluor in that row has significant spread into the dye in the column (for example PE into BV570). Areas in bright pink and red is where more attention to panel design is needed.

Spread matrix for 24 Fluors that can be use in combination

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In house unique spectra

Live/dead

DAPI

Zombie NIR

UV excited dyes

BUV496

BUV563

BUV661

BUV737

BUV805

Violet excited fluorescent protein

eBFP

eCFP

eCerulean3

Blue excited fluorescent protein

eGFP

eYFP

mVenus

ZsGreen

Red excited fluorescent protein Not optimally excited, test it before you use or wait until the 561 laser is installed

DsRed

mCherry

mPlum

mStrawberry

mRuby