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.
Code | Filter Type |
---|---|
U | Ultraviolet |
V | Violet |
BV | Blue-Violet |
B | Blue |
IB | InterferenceBlue Filter |
G | Green |
IG | Immuno GoldStaining Method |
BF | Brightfield,Phase Contrast,and DIC only |
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.
Discover how variations in excitation and barrier filters allow certain wavelengths to pass.
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 |
Protein Modification | ||||
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 |
Nuclei, Nucleic Acid | ||||
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 |
Bone | ||||
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 |
Dead Cell Staining | ||||
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 |
Nerve Cell Tracers | ||||
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 |
Membranes | ||||
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 |
Organelles | ||||
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 |
Bacteria Staining | ||||
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 |
Other Applications | ||||
NBD-Phallacidin | 465 | 530 | B, IB | Actin Fibers |
Rhodamine-Phalloidin | 550 | 580 | G, IG | Actin Fibers |
Thioflavin T | 430 | 550 | BV | Amyloid |
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