The following tables contain data about various fluorescence excitation and emission properties of over 75 commonly used fluorochromes. They are intended to provide a reference source that relates the spectral properties of these fluorochromes to commercially available filtering cubes. Table 1 is provided as a reference listing of the codes used in Table 2 for filter types.
The table below (Table 2) contains data on the peak excitation and emission wavelengths of over 75 selected fluorochromes along with suggested applications and excitation filter cube type. The selection of a wideband, superwide band or narrow band filter cube will depend on the condition of the specimen and observation purpose.
In general, a wide band filter cube is suitable in most cases. For some specimens where more brightness is desired, a superwide filter cube may be applied at some expense to contrast. If more contrast for a specific wavelength band is required for a sufficiently bright specimen, a narrow band filter cube may be appropriate.
Fluorochrome
Excitation Wavelength
Emission Wavelength
Applicable Excitation
Comment
Fluorochrome Antibody Method
Allophyco-cyanin (APC)
650
660
IY
Antibody labeling
7-Amino-4-Methyl-coumarin-3-Acetic Acid (AMCA)
350
450
U
Antibody labeling
BODIPY FL
503
512
B, IB
Antibody labeling
Cascade Blue
376
425
U
Antibody labeling
Fluorescein-Isothiocyanate (FITC)
490
520
IB
Antibody labeling
Phycoerythrin B (PE-B)
545
576
G
Antibody labeling
Phycoerythrin R (PE-R)
490, 565
578
G, IB
Antibody labeling
Rhodamine B-Iso-thiocyanate (RITC)
570
595
G
Antibody labeling
Texas Red
596
620
IY
Antibody labeling
Tetramethylrhodamine-Isothyocyanate (TRITC)
541
572
IG
Antibody labeling
8-Anilino-1-Napthalene Sulfonic
Acid Mg-salt (ANS)
385
485
V, U, BV
Hydrophobic Part Probe
Coumarin Maleimide (CMP)
385
465
U
Thiol Group
Cy 3
552
565
IG
Amino Group, Antibody Labeling
Cy 5
650
667
IY
Amino Group, Antibody Labeling
Dimethylaminonaphthalene-5-Sulfonic Acid (DANS)
380
520
U
Thiol Group
Dansyl Chloride
335
500
U
Amine Labeling
Dansyl Hydrazine
336
531
U
Carbonyl Group
Eosin-Isothiocyanate
524
548
IB
Amine Labeling
Fluorescamine
390
460
U, V
Amine Labeling
Lucifer Yellow CH
435
530
BV, B
Aldehyde, Ketone
Monochlorobimane
380
461
U
Thiol Group
o-Phtalaldehyde (OPT)
340
455
U
Amine Labeling
Resorufin
570
585
G
Thiol Group
SITS
350
420
U
Amine Labeling
Acridine Orange
490
530, 640
B, IB
Single/Double-stranded
Nuclieic Acid
Acridine Yellow
470
550
B
Nucleic Acid
Acriflavine-Feulgen
430
515
BV
DNA
Auramine O-Feulgen
460
550
B
DNA
7-Amino-Actinomycin D
555
655
IY, IG
G-C Range
Bis-Aminophenyl-Oxabizole (BAQ)-Feulgen
380
470
U
DNA
Chromomycin A3
450
570
B, BV
G-C Range
4, 6-Diamidino-2-Phenylindole HCl (DAPI)
372
456
U
A-T Range
Ethidium Bromide
545
605
G, IG
DNA•RNA
Hoechst 33258
365
465
U
A-T Range
Hoechst 33342
355
465
U
A-T Range
Mithramycin
395
570
BV
G-C Range
Olivomycin
430
545
BV
G-C Range
Pararosaniline-Feulgen
560
625
G
DNA
Propidium Iodide
530
615
IB, G, IG
DNA•RNA
Pyronin Y
540
570
IG
DNA•RNA
Quinacrine Mustard
385
525
BV
A-T Range, Q Band
Acidic Fuchsin (Pararosaniline)
540
630
G
Calcification
Calcein
495
520
IB
Bone Growth
Calcein Blue
375
435
U
Bone Growth
Tetracycline
390
560
BV, U
Calcification
Xylenol Orange
440, 570
610
G
Bone Growth
Carbonydrate Modification
Acriflavine SO2, PAS
430
515
BV
Sugar
Cell Viability Judgement & Living Cell Staining
Carboxyfluoresceindiacetate (CFDA)
495
520
IB
Esterase Substrates
Fluoresceindiacetate (FDA)
495
520
IB
Esterase Substrates
Calcein/AM
495
520
IB
Esterase Substrates
BCECF/AM
500
530
IB
Esterase Substrates
Acridine Orange
490
590
B, IB
Acid Organanelle
Ethidium Bromide
545
605
G, IB
Cell membrane Impermeability
Ethidium Homodimer
528
617
G, IB
Cell membrane Impermeability
Propidium Iodide
530
615
G, IB
Cell membrane Impermeability
Evans Blue
550
610
G
Retrograde Labeling
Fast Blue
360
410
U
Retrograde Labeling
Granular Blue
375
410
U
Retrograde Labeling
Lucifer Yellow CH
430
535
BV
Tracer
True Blue
373
404
U
Retrograde Labeling
8-Anilino-Naphthalene Sulfonic Acid (ANS)
385
485
V
Negative Ionicity
DiI
550
565
G, IG
Positive Ionicity, Tracer
DiO
484
501
B, IB
Positive Ionicity, Tracer
Dipheny-1-Hexatriene (DPH)
351
430
U
Neutral
Filipin
350
U
Cholesterol
Fluorescamine
390
460
U, V
Membrane Surface Protein
Merocyanine 540
500
572
G
Membrane Potential
Acridine Orange
490
590
B, IB
Lysosome
BODIPY FL
503
512
IB
Golgi Apparatus
DiOC6(3)
480
501
IB
Endoplasmic Reticulum
Nile Red
485-530
525-605
B, G
Lysosome
Rhodamine 123
500
540
IB
Mitochondria
Pyronin Y
540
570
IG
Mitochondria
Acridine Orange
490
530, 640
B, IB
Tubercle Bacilli
Acridine Yellow
470
550
B
Tubercle Bacilli, Acid-Fast Bacteria
Auramine O
460
550
B
Tubercle Bacilli, Acid-Fast Bacteria
Berberine Sulfate Hoechst 33258
430
550
BV
General Bacteria
Hoechst 33258
365
465
U
Mycoplasma Detection
NBD-Phallacidin
465
530
B, IB
Actin Fibers
Rhodamine-Phalloidin
550
580
G, IG
Actin Fibers
Thioflavin T
430
550
BV
Amyloid
Mortimer Abramowitz - Olympus America, Inc., Two Corporate Center Drive., Melville, New York, 11747.
Michael W. Davidson - National High Magnetic Field Laboratory, 1800 East Paul Dirac Dr., The Florida State University, Tallahassee, Florida, 32310.