Fluorochrome Data

The illumination source, filter set and detection system of a microscope dictate whether its optical system can precisely excite a given fluorochrome combination and specifically detect/discriminate different fluorochrome emissions. It is therefore important that your choice of fluorochromes reflect the specific configuration of the microscope that you will use for their visualisation. This is especially important for labelling experiments involving multiple fluorochromes, particularly in fluorochrome co-localisation analyses, and for advanced fluorescence microscopy techniques. Helpful advice on fluorochrome selection can be found here.                                                                                                         

Wavelengths of visible light

Colour ~ Wavelength interval (nm)
Violet 380-430
Blue 430-500
Cyan 500-520
Green 520-565
Yellow 565-590
Orange 590-625
Red 625-740

 

Spectral Properties of Common Flurochromes

Fluorochrome Ex (max) Em (max)
Acridine Orange 500 (+DNA)460 (+RNA) 526 (+DNA)650 (+RNA)
Alexa 405 402 421
Alexa 430 434 539
Alexa 488 495 519
Alexa 500 503 525
Alexa 514 518 540
Alexa 532 531 554
Alexa 546 556 573
Alexa 555 555 565
Alexa 568 578 603
Alexa 594 590 617
Alexa 610 612 628
Alexa 633 632 647
Alexa 635 633 647
BCECF 482 (low pH)503 (high pH) 520 (low pH)528 (high pH)
BOBO, BOPRO 462 481
BODIPY FL 505 513
BODIPY TMR-X 542 574
BODIPY TR-X 589 617
Calcein 494 517
6-Carboxyrhodamine 6G 525 555
Cascade Blue 400 420
Cascade Yellow 402 545
DAPI 358 461
DiI 549 565
DsRed (OFP) 558 583
EBFP 383 445
ECFP 439 476
EGFP 484 507
EYFP 514 527
Eosin 524 544
ER-Tracker 372 556
Ethidium Bromide 518 605
Fluo-3 506 526
Fluo-4 494 516
Fluoroscein (FITC) 494 518
Fura-2 363 (low Ca2+)335 (high Ca2+) 512 (low Ca2+)505 (high Ca2+)
Fura Red 472 (low Ca2+)436 (high Ca2+) 657 (low Ca2+)637 (high Ca2+)
GFP 395/475 509
Hoechst 33258, 33342 352 461
JC-1 515 529/590
Lissamine Rhodamine B 570 590
Lucifer Yellow CH 428 536
LysoTracker Green 504 511
mApple (RFP) 568 592
mBanana (YFP) 540 553
mCherry (RFP) 587 610
mOrange (OFP) 584 562
mPlum (RFP) 590 637
mRasberry (RFP) 598 625
mTangerine (OFP) 568 585
MitoTracker Green 490 516
Nile Red 549 628
Oregon Green 488 496 524
Oregon Green 514 511 530
Pacific Blue 410 455
Pacific Green 410 500
Pacific Orange 410 500
POPO-1, PO-PRO-1 434 456
Propidium Iodide 536 617
Rhodamine Red 570 590
R-phycoerythrin 565 575
Rhodamine 123 507 529
SYTO9 483 503
SYTOX Blue 445 470
SYTO Green 504 523
SYTO Orange 547 570
TRITC 555 580
Texas Red 595 615
TOTO-1, TO-PRO-1 514 533
TOTO-3, TO-PRO-3 642 660
YOYO-1, YO-PRO-1 491 509
YOYO-3, YO-PRO-3 612 631

 

Fluorescence spectral viewer (via Molecular probes)

This tool is useful for the analysis of excitation and emission spectra of commonly used fluorochromes, particularly for multiplexed fluorescent labelling studies.

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