The fluorescence filters listed are available for Infinite 200 PRO and Spark multimode readers with filter-based fluorescence optics.
The fluorescence filters listed are available for Infinite 200 PRO and Spark multimode readers with filter-based fluorescence optics.
The recommended filter combinations for each dye were selected according to the excitation and
emission properties (1) of the dye, and based on a formula to to avoid crosstalk (3). Please be aware that excitation and emission properties of a fluorescent dye will vary with assay conditions – such as pH, salt concentrations and labeling partners – and that derivatives of a dye may have altered spectra. Suitability of the filter recommendation for your specific assay cannot be guaranteed, and other filter combinations might also be suitable. Users must always independently evaluate and validate filters for their intended use.
Please download the Filter overview here:
Filter overview for fluorescent dyes Infinite® 200 PRO and Spark®
| Dye | Dye properties | Excitation filter | Emission filter | |
| Ex | Ex | WL (BW) | WL (BW) | |
| 5´-ROX (5-carboxy-X-rhodamine) | 578 nm | 604 nm | 560 nm (20 nm) | 610 nm (20 nm) |
| 5-FAM (5-Carboxyfluorescein) | 492 nm | 518 nm | 485 nm (20 nm) | 535 nm (25 nm) |
| alamarBlue® (resazurin/resurofin) | 571 nm | 583 nm | 560 nm (10 nm) | 590 nm (20 nm) |
| Alexa Fluor® 532 | 531 nm | 554 nm | 520 nm (10 nm) | 560 nm (20 nm) |
| Alexa Fluor 546 | 556 nm | 573 nm | 540 nm (25 nm) | 590 nm (20 nm) |
| Alexa Fluor 555 | 555 nm | 565 nm | 530 nm (25 nm) | 595 nm (35 nm) |
| Alexa Fluor 647 | 650 nm | 668 nm | 620 nm (20 nm) | 680 nm (30 nm) |
| Allophycocyanin (APC) | 645 nm | 660 nm | 620 nm (20 nm) | 670 nm (25 nm) |
| AMC (amino-methylcoumarin), pH 9 | 360 nm | 448 nm | 380 nm (20 nm) | 430 nm (20 nm) 460 nm (20 nm) |
| Amplex® Red (resorufin) | 571 nm | 585 nm | 540 nm (25 nm) | 590 nm (20 nm) |
| BCECF | 503 nm | 528 nm | 485 nm (20 nm) | 535 nm (25 nm) |
| Bodipy 581/591 | 584 nm | 592 nm | 560 nm (20 nm) | 625 nm (35 nm) |
| Bodipy FI | 505 nm | 513 nm | 485 nm (20 nm) | 535 nm (25 nm) |
| Bodipy R6G SE | 528 nm | 547 nm | 505 nm (20 nm) | 560 nm (20 nm) |
| Bodipy TMR | 542 nm | 570 nm | 530 nm (25 nm) | 580 nm (20 nm) 595 nm (35 nm) |
| Bodipy TR | 589 nm | 617 nm | 560 nm (20 nm) | 635 nm (35 nm) |
| Calcein AM | 494 nm | 517 nm | 485 nm (20 nm) | 535 nm (25 nm) |
| Cascade Blue | 400 nm | 420 nm | 380 nm (20 nm) | 430 nm (20 nm) |
| CFP | 435 nm | 477 nm | 430 nm (20 nm) | 485 nm (20 nm) |
| Cy3 | 554 nm | 566 nm | 530 nm (25 nm) | 580 nm (20 nm) 595 nm (35 nm) |
| Cy3.5 | 576 nm | 598 nm | 560 nm (20 nm) | 620 nm (20 nm) 635 nm (35 nm) |
| Cy5 | 647 nm | 665 nm | 620 nm (20 nm) | 680 nm (30 nm) |
| Cy5.5 | 672 nm | 692 nm | 635 nm (35 nm) | 720 nm (40 nm) |
| DAPI (DNA) | 358 nm | 461 nm | 360 nm (35 nm) | 465 nm (35 nm) |
| DCFDA | 505 nm | 525 nm | 485 nm (20 nm) | 535 nm (25 nm) 540 nm (25 nm) |
| DELFIA® (europium) | 340 nm (35 nm) | 612 nm (10 nm) | ||
| DsRed | 556 nm | 583 nm | 530 nm (25nm) | 590 nm (20 nm) 595 nm (35 nm) |
| DsRed | 556 nm | 583 nm | 540 nm (25 nm) | 590 nm (20 nm) |
| eGFP | 489 nm | 511 nm | 485 nm (20 nm) | 535 nm (25 nm) |
| Ethidium homodimer-1 (EthD-1) | 528 nm | 617 nm | 535 nm (25 nm) | 620 nm (20 nm) 635 nm (35 nm) |
| Europium – chelate | 365 nm | 610 nm | 340 nm (20 nm) 340 nm (35 nm) |
610 nm (20 nm) |
| eYFP | 514 nm | 527 nm | 485 nm (20 nm) | 535 nm (25 nm) |
| eYFP | 514 nm | 527 nm | 510 nm (10 nm) | 535 nm (10 nm) |
| FDA (fluoresceindiacetat) | 498 nm | 516 nm | 485 nm (20 nm) | 535 nm (25 nm) |
| FITC (fluorescein-5-isothiocyanate) | 495 nm | 519 nm | 485 nm (20 nm) | 535 nm (25 nm) |
| Fluo-3 | 506 nm | 527 nm | 485 nm (20 nm) 495nm (10 nm) |
520nm (10 nm) 535nm (10 nm) |
| Fluorescein | 489 nm | 517 nm | 485 nm (20 nm) | 535nm (25 nm) |
| Fura-2 (Ex changes from 366 nm to 336 nm on Ca2+ binding) |
366 nm (336 nm) |
505 nm | 340 nm (20 nm) 380 nm (20 nm) |
510 nm (25 nm) |
| Hoechst® 33258 (DNA) | 352 nm | 455 nm | 360 nm (35 nm) | 465 nm (35 nm) |
| Hoechst 33342 | 348 nm | 455 nm | 360 nm (35 nm) | 465 nm (35 nm) |
| HTRF® (Cisbio, europium) both Em filter necessary | 320 nm (25 nm) | 620 nm (10 nm) 665 nm (8 nm) |
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| HTRF (Cisbio, Terbium) both Em filter necessary |
340 nm (35 nm) | 620 nm (10 nm) 665 nm (8 nm) |
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| Lissamine™ Rhodamine | 570 nm | 590 nm | 560 nm (10 nm) 560 nm (20 nm) |
610 nm (20 nm) |
| mCherry | 587 nm | 610 nm | 560 nm (20 nm) | 620 nm (20 nm) |
| Monobromobimane (mBBr) | 394 nm | 490 nm | 380 nm (20 nm) 400 nm (20 nm) |
465 nm (35 nm) 485 nm (20 nm) |
| NADH | 340 nm | 466 nm | 340 nm (20 nm) | 465 nm (35 nm) |
| NanoOrange® | 470 nm | 570 nm | 465 nm (35 nm) 485 nm (20 nm) |
560 nm (20 nm) 580 nm (20 nm) 580 nm (30 nm) |
| Phycoerythrin B | 545 nm | 575 nm | 535 nm (10 nm) | 580 nm (20 nm) 590 nm (20 nm) |
| Phycoerythrin R | 565 nm | 576 nm | 540 nm (25 nm) | 590 nm (20 nm) |
| PicoGreen® (DNA) | 502 nm | 522 nm | 485 nm (20 nm) | 535 nm (25 nm) |
| PO-PRO-1 | 280 nm | 457 nm | 280 nm (15 nm) | 460 nm (10 nm) 460 nm (20 nm) |
| Propidium iodide (PI) | 535 nm | 617 nm | 535 nm (25 nm) | 620 nm (20 nm) |
| RFP | 555 nm | 584 nm | 535 nm (25 nm) | 590 nm (20 nm) |
| SYBR® Green I (DNA) | 498 nm | 522 nm | 485 nm (20 nm) | 535 nm (25 nm) |
| SYTO™ 17 (DNA) | 620 nm | 638 nm | 610 nm (20 nm) | 670 nm (25 nm) |
| TAMRA | 553 nm | 576 nm | 535 nm (25 nm) | 590 nm (10 nm) 590 nm (20 nm) |
| Terbium – chelate (TR-FRET; LanthaScreen™, Invitrogen) |
340 nm (35nm) | 495 nm (10 nm) | ||
| Texas Red | 592 nm | 614 nm | 560 nm (20 nm) | 620 nm (20 nm) 635 nm (35 nm) |
| Texas Red | 592 nm | 614 nm | 580 nm (20 nm) | 620 nm (10 nm) |
| TMR (Tetramethylrhodamine) | 552 nm | 578 nm | 535 nm (25 nm) | 590 nm (20 nm) |
| TRITC | 555 nm | 576 nm | 535 nm (25 nm) 540 nm (25 nm) |
590 nm (20 nm) |
| Tryptophan | 278 nm | 354 nm | 280 nm (15 nm) | Scanning (300 - 500 nm) for conformation studies |
| YFP | 513 nm | 530 nm | 485 nm (20 nm) 495 nm (10 nm) |
535 nm (25 nm) 540 nm (25 nm) |
How to select a suitable filter pair for a fluorescent dye
1) Obtain the excitation and emission maxima from the dye’s data sheet, suitable literature or database (eg. https://www.nightsea.com/sfa-sharing/fluorescence-spectra-viewers*).
2) Select filters with wavelength properties (WL + BW) covering the excitation or emission maxima.
3) Check the filter selection for spectral overlap using the following formula:
WL EM - BW EM - WL EX - BW EX = ≥5 nm
WL = Wavelength of the filter EX = Excitation
BW = Bandwidth of the filter EM = Emission
At least 5 nm distance is recommended between the upper end of the spectral range of the excitation filter (WL EX + BW EX) and the lower end of the spectral range of the emission filter (WL EM - BW EM).
4) Evaluate and validate the selected filters for your specific assay and use.
If no suitable filters are available in the list, please contact your local Tecan sales representative.
Example: fluorescein
· Excitation maximum: 489 nm
· Emission maximum: 517 nm
· Recommended excitation filter: 485 nm (20 nm)
· Recommended emission filter: 535 nm (25 nm)
535 - 25 - 485 - 20 = ≥5 nm
The recommended filters for fluorescein cover the respective maxima (excitation and emission),
and show the recommended minimal spectral distance of 5 nm.
Important information:
Instrument measurement specifications are tested and validated for specific dyes and filters only, as stated in the instructions for use for the instrument. Measurement limits may be different for other dyes and filters. Users must always independently evaluate and validate the suitability of filters for their intended use.
* Please note that Tecan is not responsible for the content of this webpage.
Further Help
For further help, please refer to our Helpdesk.
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