Fluorochrome excitation with one-photon and two-photon systems.
(Updated 09-08-2003)
Some of the information provided (e.g. 2P exc with ">" and "<" marks) are from Leica. Information provided by Leica is based a Leica system with a SpectraPhysics laser. Other info is empirical on our Leica SP2 with a Coherent Mira 9000 system or provided by another source. If you have more fluorochromes to add or wish to modify the existing data, please contact us with the dye name and any synonyms, and other information that will make this chart more useful to all users. jshields@cb.uga.edu or farmer@cb.uga.edu
Application |
Dye |
Solvents |
1P exc |
2P exc |
UV exc |
Max em |
Cell wall/ fibers |
Aniline blue |
water |
500 |
790 |
607-610 |
|
|
Calcofluor |
water |
440/500-520 |
780>820 |
340-400 |
400-440 |
(Polysciences) |
FungiFluor (cellufluor) |
water |
350-400 |
780 |
250-400 |
350-400 |
|
|
|
|
|
|
|
Nucleic acid stains |
DAPI, Hoechst |
Water |
350/470 |
700, 780>820 |
345 |
455, 478 |
|
Feulgen |
Water, ethanol |
480/560 |
780>820 |
460 |
550 |
|
Propidium Iodide (PI) |
Water |
540 |
770-790 |
305, 536 |
617 |
|
Acridine orange |
|
467 |
|
503 |
530, 640 |
|
Ethidium Bromide |
Water |
488 |
|
493 |
620 |
(Molecular Probes) |
SYTO 13 |
|
|
|
488 |
509 |
|
SYTO 85 |
|
|
|
579 |
599 |
|
YOYO-1 |
|
|
|
491 |
508 |
|
|
|
|
|
|
|
Protein conjugates |
FITC |
Ethanol,Water |
490/525 |
780>820 |
495 |
519 |
|
*FITC-dextran (fluoro-emerald) |
|
494 |
830 |
|
521 |
|
TRITC (rhodamine) |
|
541/572 |
800-840 |
547 |
572 |
|
*TRITC-dextran (fluoro-ruby) |
|
555 |
850 |
|
580 |
|
Rhodamine B |
|
|
840 |
|
600 |
|
Texas red |
|
596/620 |
780 |
589 |
515 |
|
*Texas Red-dextran |
|
595 |
880 |
|
516 |
(Jackson Imm. Res.) |
CY2 |
water |
489/506 |
780>800 |
489 |
506 |
|
CY3 |
water |
550/570 |
780 |
512, 552 |
565, 615 |
|
CY5 |
water |
649/670 |
780<820 |
625-650 |
670 |
|
AMCA (7 amino-4 methylcoumarin-3-acetate) |
|
431/498 |
780 |
347 |
444 |
(Molecular Probes) |
*Alexa 488 |
|
495 |
830 |
495 |
519 |
|
Alexa 532 |
|
|
|
531 |
554 |
|
Alexa 546 |
|
|
|
556 |
573 |
|
Alexa 568 |
|
|
|
578 |
603 |
|
*Alexa 594 |
|
590 |
840 |
590 |
617 |
|
Oregon Green |
|
488 |
800-820 |
488 |
520 |
|
|
|
|
|
|
|
Gene expression |
BFP |
|
395/509 |
780>820 |
382 |
448 |
(ClonTech) |
CFP |
|
434/477 |
780>840 |
430 |
476 |
|
GFP |
|
488/507 |
800-850 |
498 |
516 |
|
E GFP |
|
488 |
900-950 |
498 |
507 |
|
YFP |
|
514/527 |
860<900 |
520 |
532 |
|
*DsRed1 & 2 (redFP) |
|
558 |
920 |
558 |
583 |
|
|
|
|
|
|
|
Mitochondria |
Rhodamine 123 |
|
507/529 |
780-860 |
480 |
550 |
(Molecular Probes) |
Mito Tracker red |
DMSO |
|
|
579 |
599 |
|
MitoTracker red 594 |
|
|
|
598 |
630 |
|
MitoTracker Green FM |
|
|
|
490 |
516 |
|
|
|
|
|
|
|
|
|
|
|
|
|
|
Lipids |
Nile Red |
ethanol |
|
810 |
559 |
640 |
|
|
|
|
|
|
|
Calcium |
Calcium green |
|
488/530 |
780>820 |
506 |
531 |
|
Ca Green/
Texas Red |
|
488/530, 596/620 |
780 |
|
|
|
Yellow Chameleon |
|
464/527 |
780>820 |
|
|
|
Indo1-free |
|
|
700 |
|
490 |
|
Indo1-Ca++ |
|
|
590 |
|
405 |
|
Fura-2 free |
|
|
700 |
|
362 |
|
Fura-2 Ca++ |
|
|
? |
|
? |
|
|
|
|
|
|
|
Viability, esterases |
Fluorescein Diacetate |
acetone |
495/520 |
780>820 |
490-512 |
515-535 |
(PolySciences) |
CTC (viab – bacteria) |
|
|
720-740 |
450 |
630 |
|
|
|
|
|
|
|
Neuronal tracer |
*DiD |
|
644 |
830 |
|
665 |
|
Lucifer Yellow CH |
EtOH (water) |
|
|
488 (428) |
531 |
|
|
|
|
|
|
|
Lysosomes |
Lysotracker (Mol. Probes) |
MeOH, DMSO |
|
|
577 |
590 |
|
|
|
|
|
|
|
Membrane Probes |
*DiI |
|
549 |
900 |
561 |
565 |
|
*DiA |
|
456 |
880 |
|
590 |
|
*DiO |
|
|
|
|
|
|
*Di-8-ANEPPS |
|
467 |
880 |
|
631 |
|
Filipin |
|
|
|
380 |
510 |
|
|
|
|
|
|
|
triple probe (on Leica/Coherent
system) |
Dapi, FITC, and Rhodamine |
|
|
740 |
|
see above for each fluor. |
* From Ruthhazer & Cline
