荧光滤光片是应用于生物医学和生命科学仪器的关键元件,主要作用是在生物医学荧光检验分析系统中分离和选择物质的激发光与发射荧光的特征波段光谱。荧光滤光片一般包含三片组合,即激发滤光片、二色镜和发射滤光片。
激发滤光片(Exciting Filter, Exciter Filter,Excitation Filter ):在荧光显微镜中,只有激发荧光的波长可通过的滤光片。也有直接使用激光作为激发光。
二色镜(Dichroic Mirror,Dichroic Beamsplitter,Dichromatic Beamsplitter):又称二向色滤光片或分色镜。与显微镜的光路呈45°角放置。这片滤光片反射一种颜色光(激发光)并且透射另一种颜色光(发射光),激发光的反射率大于90%,发射光的透射率大于90%。光谱中不能透过的部分被反射而不是被吸收。滤光片在透射光和反射光下的颜色互为补色,因而又称二向色滤光片。
发射滤光片(Emitting Filter,Emission Filter ,Barrier Filter,Emitter):选择并传输样本发出的荧光,其他范围光线截止。发射光的波长比激发光的波长要长。可以选择带通滤光片或者长波通滤光片作为发射滤光片。
兆九光电生产的PCR仪滤光片一般常用的有4通道,有时是5通道,分别是:不同的厂家,波长位置略有差异。截止深度为OD6以上,截止范围一般为200-800nm, 或200-900nm
下表是一些常用荧光染料所对应使用的荧光滤光片组合,如有需要欢迎来电咨询:
荧光染料 |
激发波长 峰值 |
发射波长 峰值 |
激发滤光片 |
发射滤光片 |
二色镜 |
|||
CWL |
FWHM |
CWL |
FWHM |
高反射范围 |
高透过范围 |
|||
Alexa Fluor® 350 | 342 |
442 |
350 |
50 |
460 |
50 |
330-380 | 430-700 |
Alexa Fluor® 488 | 499 |
520 |
480 |
40 |
535 |
50 |
400-500 | 520-700 |
Alexa Fluor® 514 | 518 |
543 |
500 |
40 |
560 |
50 |
450-510 | 530-650 |
Alexa Fluor® 532 | 531 |
554 |
530 |
30 |
575 |
40 |
450-540 | 560-650 |
Alexa Fluor® 546 | 556 |
572 |
545 |
25 |
590 |
30 |
450-540 | 570-700 |
Alexa Fluor® 555 | 553 |
565 |
545 |
25 |
590 |
30 |
450-540 | 570-700 |
Alexa Fluor® 568 | 579 |
603 |
570 |
25 |
615 |
40 |
500-580 | 600-700 |
Alexa Fluor® 594 | 590 |
618 |
580 |
25 |
625 |
30 |
500-590 | 610-700 |
Alexa Fluor® 633 | 631 |
647 |
605 |
50 |
670 |
50 |
550-620 | 645-750 |
Alexa Fluor® 647 | 653 |
668 |
640 |
30 |
690 |
50 |
600-650 | 665-730 |
Alexa Fluor® 680 | 679 |
702 |
650 |
45 |
720 |
60 |
600-660 | 690-800 |
Alexa Fluor® 700 | 696 |
719 |
650 |
45 |
720 |
60 |
600-660 | 690-800 |
Alexa Fluor® 750 | 751 |
776 |
710 |
75 |
810 |
90 |
600-730 | 770-900 |
AmCyan | 458 |
489 |
435 |
20 |
480 |
40 |
425-445 | 460-600 |
APC (Allophycocyanin) | 650 |
660 |
620 |
60 |
700 |
75 |
600-650 | 665-730 |
AsRed 2 | 576 |
592 |
545 |
30 |
620 |
60 |
530-560 | 580-700 |
ATTO 390 | 392 |
477 |
395 |
25 |
460 |
50 |
350-410 | 430-550 |
ATTO 425 | 436 |
484 |
445 |
30 |
500 |
40 |
400-460 | 480-600 |
ATTO 465 | 453 |
507 |
470 |
40 |
525 |
50 |
400-480 | 500-600 |
ATTO 488 | 500 |
525 |
480 |
40 |
535 |
50 |
400-500 | 520-600 |
ATTO 550 | 553 |
576 |
545 |
20 |
590 |
30 |
450-550 | 570-700 |
ATTO 633 | 629 |
658 |
605 |
50 |
670 |
50 |
550-620 | 640-750 |
ATTO 647N | 645 |
668 |
620 |
60 |
700 |
75 |
590-650 | 665-735 |
ATTO 680 | 680 |
700 |
650 |
45 |
720 |
60 |
600-670 | 690-800 |
BCECF (pH 5.2) | 482 |
520 |
480 |
40 |
540 |
40 |
350-500 | 520-650 |
BCECF (pH 9.0) | 502 |
528 |
480 |
40 |
540 |
40 |
350-500 | 520-650 |
Biosearch Blue | 352 |
447 |
350 |
50 |
460 |
50 |
330-390 | 430-550 |
BODIPY FL/pH7.2 | 505 |
512 |
480 |
40 |
535 |
50 |
400-500 | 520-600 |
Brilliant Violet™ 421 | 408 |
422 |
395 |
25 |
430 |
30 |
350-405 | 415-500 |
Brilliant Violet™ 480 | 437 |
478 |
435 |
20 |
480 |
30 |
425-445 | 460-600 |
CAL Fluor® Gold 540 | 522 |
544 |
530 |
30 |
575 |
40 |
450-540 | 555-650 |
CAL Fluor® Orange 560 | 538 |
559 |
530 |
30 |
575 |
40 |
450-540 | 555-650 |
CAL Fluor® Red 590 | 569 |
591 |
570 |
25 |
615 |
40 |
500-580 | 595-700 |
CAL Fluor® Red 610 | 590 |
610 |
580 |
25 |
625 |
30 |
500-590 | 605-700 |
CAL Fluor® Red 635 | 618 |
637 |
605 |
50 |
670 |
50 |
550-620 | 645-750 |
Calcein | 494 |
517 |
480 |
40 |
535 |
50 |
400-500 | 515-600 |
Calcium Green™-1 | 507 |
529 |
480 |
40 |
535 |
50 |
400-500 | 515-600 |
Cerulean | 434 |
473 |
435 |
20 |
480 |
40 |
425-445 | 460-600 |
CFP | 433 |
475 |
435 |
20 |
480 |
40 |
425-445 | 460-600 |
Citrine | 514 |
529 |
500 |
20 |
535 |
30 |
480-505 | 520-650 |
Coumarin | 360 |
448 |
350 |
50 |
460 |
50 |
330-390 | 430-550 |
Cy2™ | 492 |
507 |
480 |
40 |
535 |
50 |
400-500 | 515-600 |
Cy3.5™ | 578 |
591 |
570 |
25 |
615 |
40 |
500-580 | 595-700 |
Cy3™ | 554 |
566 |
545 |
25 |
605 |
70 |
450-550 | 570-700 |
Cy5.5™ | 675 |
695 |
650 |
45 |
720 |
60 |
600-660 | 690-800 |
Cy5™ | 649 |
666 |
620 |
60 |
700 |
75 |
590-650 | 665-735 |
Cy7™ | 757 |
781 |
710 |
75 |
810 |
90 |
600-730 | 770-900 |
DAPI | 359 |
461 |
350 |
50 |
460 |
50 |
330-390 | 430-550 |
Di-8-ANEPPS non-ratiometric | 466 |
630 |
480 |
30 |
620 |
60 |
400-495 | 515-700 |
DiA | 457 |
586 |
480 |
30 |
620 |
60 |
400-495 | 515-700 |
DiD | 648 |
670 |
620 |
60 |
700 |
75 |
590-650 | 665-735 |
DiI | 551 |
569 |
545 |
25 |
605 |
70 |
450-550 | 570-700 |
DiO | 490 |
506 |
470 |
40 |
525 |
50 |
400-480 | 500-600 |
DiR | 750 |
782 |
710 |
75 |
810 |
90 |
600-730 | 770-900 |
Draq5 | 647 |
683 |
620 |
60 |
700 |
75 |
590-650 | 665-735 |
DsRed | 559 |
583 |
545 |
25 |
605 |
70 |
450-550 | 570-700 |
DyLight 350 | 351 |
434 |
350 |
50 |
460 |
50 |
330-390 | 430-550 |
DyLight 405 | 398 |
420 |
405 |
20 |
460 |
50 |
350-410 | 430-550 |
DyLight 549 | 553 |
569 |
545 |
25 |
605 |
70 |
450-550 | 570-700 |
DyLight 594 | 593 |
618 |
580 |
25 |
625 |
30 |
500-590 | 605-700 |
DyLight 633 | 620 |
649 |
605 |
50 |
670 |
50 |
550-620 | 645-750 |
DyLight 649 | 651 |
673 |
620 |
60 |
700 |
75 |
590-650 | 665-735 |
DyLight 680 | 676 |
705 |
650 |
45 |
720 |
60 |
600-660 | 690-800 |
DyLight 750 | 751 |
778 |
710 |
75 |
810 |
90 |
600-730 | 770-900 |
DyLight 800 | 770 |
795 |
740 |
40 |
800 |
40 |
650-750 | 780-900 |
EBFP2 | 385 |
448 |
395 |
25 |
460 |
50 |
350-410 | 430-550 |
ECFP | 434 |
477 |
435 |
20 |
480 |
40 |
425-445 | 460-600 |
EGFP | 488 |
507 |
470 |
40 |
525 |
50 |
400-480 | 500-600 |
Emerald GFP | 489 |
510 |
470 |
40 |
525 |
50 |
400-480 | 500-600 |
Eosin | 525 |
546 |
500 |
40 |
560 |
50 |
450-510 | 530-650 |
Ethidium bromide | 518 |
603 |
545 |
30 |
620 |
60 |
530-560 | 580-700 |
Ethidium homidimer-1/DNA | 527 |
617 |
545 |
30 |
620 |
60 |
530-560 | 580-700 |
EYFP/pH 7 | 514 |
527 |
500 |
20 |
535 |
30 |
480-505 | 520-650 |
FAM | 492 |
518 |
480 |
40 |
535 |
50 |
400-500 | 515-600 |
FITC | 495 |
519 |
480 |
40 |
535 |
50 |
400-500 | 515-600 |
FlAsH-CCPFCC | 511 |
530 |
500 |
20 |
535 |
30 |
480-505 | 520-650 |
Fluo-3 | 506 |
527 |
500 |
20 |
535 |
30 |
480-505 | 520-650 |
Fluo-4 | 494 |
516 |
480 |
40 |
535 |
50 |
400-500 | 515-600 |
FM™ 1-43 | 473 |
579 |
470 |
30 |
580 |
60 |
400-495 | 515-700 |
FM™ 4-64 | 508 |
730 |
510 |
40 |
730 |
100 |
450-590 | 605-800 |
Fura-2/Ca2+ - free | 366 |
515 |
365 |
50 |
510 |
80 |
330-390 | 430-700 |
Fura-2/Ca2+ - saturated | 336 |
505 |
335 |
30 |
510 |
80 |
320-390 | 430-700 |
FusionRed | 577 |
604 |
560 |
40 |
630 |
75 |
500-570 | 595-750 |
GFP | 489 |
511 |
470 |
40 |
525 |
50 |
400-480 | 500-600 |
HEX,SE | 533 |
559 |
530 |
30 |
575 |
40 |
450-540 | 555-700 |
Hoechst 33258 | 352 |
455 |
350 |
50 |
460 |
50 |
330-390 | 430-700 |
Indo-1/Ca2+ -free | 346 |
480 |
350 |
50 |
460 |
50 |
330-390 | 430-700 |
Indo-1/Ca2+ -saturated | 330 |
400 |
350 |
40 |
420 |
40 |
310-380 | 395-550 |
JOE | 520 |
548 |
500 |
40 |
560 |
50 |
450-510 | 530-650 |
Killer Red | 585 |
611 |
560 |
40 |
630 |
75 |
500-570 | 595-750 |
Li-Cor IRDye® 680LT | 674 |
693 |
650 |
45 |
720 |
60 |
600-660 | 690-800 |
Li-Cor IRDye® 800CW | 778 |
794 |
740 |
40 |
800 |
40 |
650-750 | 780-900 |
Lucifer yellow | 428 |
544 |
420 |
40 |
535 |
40 |
390-445 | 460-600 |
LysoTracker Blue | 373 |
422 |
395 |
25 |
460 |
50 |
350-410 | 430-550 |
LysoTracker Green | 501 |
509 |
470 |
40 |
525 |
50 |
400-480 | 500-600 |
LysoTracker Red | 573 |
592 |
560 |
40 |
630 |
75 |
500-570 | 595-750 |
LysoTracker Yellow HCK-123 | 488 |
565 |
500 |
40 |
560 |
50 |
450-510 | 530-700 |
mCherry | 587 |
610 |
560 |
40 |
630 |
75 |
500-570 | 595-750 |
mCitrine | 514 |
529 |
500 |
20 |
535 |
30 |
480-505 | 520-650 |
MitoTracker™ Deep Red | 641 |
662 |
620 |
60 |
700 |
75 |
590-650 | 665-735 |
MitoTracker™ Green | 490 |
512 |
470 |
40 |
525 |
50 |
400-480 | 500-600 |
MitoTracker™ Orange | 551 |
575 |
550 |
20 |
590 |
30 |
450-550 | 570-700 |
MitoTracker™ Red | 578 |
598 |
560 |
40 |
630 |
75 |
500-570 | 595-750 |
mKate2 | 588 |
633 |
560 |
40 |
630 |
75 |
500-570 | 595-750 |
mKO | 549 |
559 |
530 |
30 |
575 |
40 |
450-540 | 555-700 |
mOrange2 | 551 |
565 |
530 |
30 |
575 |
40 |
450-540 | 555-700 |
mPlum | 589 |
649 |
560 |
40 |
650 |
40 |
450-590 | 605-800 |
mRFP1 | 590 |
612 |
560 |
40 |
630 |
75 |
500-570 | 595-750 |
mTFP1 | 462 |
492 |
445 |
30 |
500 |
40 |
400-460 | 480-600 |
mWasabi | 493 |
509 |
470 |
40 |
525 |
50 |
400-480 | 500-600 |
NBD-X (MeOH) | 467 |
538 |
480 |
40 |
535 |
50 |
400-500 | 515-700 |
Nile blue | 631 |
660 |
620 |
60 |
700 |
75 |
590-650 | 665-735 |
NirFP | 604 |
669 |
600 |
40 |
670 |
40 |
580-620 | 650-800 |
Oregon Green™ 488 | 498 |
526 |
480 |
40 |
535 |
50 |
400-500 | 515-700 |
Oregon Green™ 514 | 513 |
532 |
500 |
20 |
535 |
30 |
480-505 | 520-650 |
Pacific Blue™ | 404 |
455 |
405 |
20 |
460 |
50 |
350-410 | 430-550 |
Propidium Iodide (PI) | 305 |
617 |
545 |
30 |
620 |
60 |
530-560 | 580-700 |
Pulsar™ 650 | 460 |
650 |
440 |
40 |
630 |
75 |
400-480 | 500-600 |
Qdot® 525 Nanocrystals | 300 |
525 |
425 |
50 |
525 |
30 |
370-470 | 490-700 |
Qdot® 545 Nanocrystals | 300 |
543 |
425 |
50 |
545 |
30 |
370-470 | 490-700 |
Qdot® 565 Nanocrystals | 300 |
564 |
425 |
50 |
565 |
30 |
370-470 | 490-700 |
Qdot® 585 Nanocrystals | 300 |
588 |
425 |
50 |
585 |
30 |
370-470 | 490-700 |
Qdot® 605 Nanocrystals | 300 |
602 |
425 |
50 |
605 |
30 |
370-470 | 490-700 |
Qdot® 625 Nanocrystals | 300 |
621 |
425 |
50 |
625 |
30 |
370-470 | 490-700 |
Qdot® 655 Nanocrystals | 300 |
654 |
425 |
50 |
655 |
30 |
370-470 | 490-700 |
Qdot® 705 Nanocrystals | 300 |
702 |
425 |
50 |
705 |
30 |
370-470 | 490-700 |
Qdot® 800 Nanocrystals | 300 |
792 |
425 |
50 |
800 |
40 |
370-470 | 490-700 |
Quasar® 570 | 548 |
566 |
530 |
30 |
575 |
40 |
450-540 | 555-700 |
Quasar® 670 | 647 |
670 |
620 |
60 |
700 |
75 |
590-650 | 665-735 |
Quasar® 705 | 690 |
750 |
650 |
45 |
720 |
60 |
600-660 | 690-800 |
ReAsH-CCPGCC | 592 |
606 |
560 |
40 |
630 |
75 |
500-570 | 595-750 |
Resorufin | 571 |
584 |
545 |
20 |
590 |
30 |
450-550 | 570-700 |
Rhod-2 | 553 |
577 |
545 |
25 |
605 |
70 |
450-550 | 570-700 |
Rhodamine 123 | 507 |
529 |
500 |
20 |
535 |
30 |
480-505 | 520-650 |
Rhodamine 6G | 525 |
547 |
500 |
40 |
560 |
50 |
450-510 | 530-700 |
Rhodamine Red-X | 573 |
591 |
560 |
40 |
630 |
75 |
500-570 | 595-750 |
ROX | 578 |
604 |
560 |
40 |
630 |
75 |
500-570 | 595-750 |
R-phycoerythrin | 565 |
578 |
545 |
10 |
585 |
40 |
500-550 | 570-800 |
SBFI/Na+ -free | 340 |
560 |
350 |
40 |
560 |
100 |
330-390 | 430-700 |
SBFI/Na+ -saturated | 337 |
527 |
350 |
40 |
560 |
100 |
330-390 | 430-700 |
Sulforhodamine 101-EtOH | 578 |
593 |
560 |
40 |
630 |
75 |
500-570 | 595-750 |
SYBR® Green I | 498 |
522 |
480 |
40 |
535 |
50 |
400-500 | 515-700 |
SYTO 60 | 650 |
681 |
620 |
60 |
700 |
75 |
590-650 | 665-735 |
SYTO 9 | 483 |
500 |
470 |
40 |
525 |
50 |
400-480 | 500-600 |
TagBFP | 402 |
457 |
405 |
20 |
460 |
50 |
350-410 | 430-550 |
TagRFP | 556 |
584 |
545 |
25 |
605 |
70 |
450-550 | 570-700 |
TAMRA | 555 |
580 |
545 |
25 |
605 |
70 |
450-550 | 570-700 |
tdTomato | 554 |
581 |
545 |
25 |
605 |
70 |
450-550 | 570-700 |
TET | 521 |
542 |
500 |
40 |
560 |
50 |
450-510 | 530-700 |
Tetracycline | 405 |
550 |
405 |
40 |
550 |
60 |
350-460 | 480-650 |
Tetramethylrhodamine dextran | 554 |
582 |
545 |
25 |
605 |
70 |
450-550 | 570-700 |
Texas Red® | 592 |
614 |
560 |
40 |
630 |
75 |
500-570 | 595-750 |
TO-PRO™-3 | 642 |
657 |
620 |
60 |
700 |
75 |
590-650 | 665-735 |
TRITC | 552 |
578 |
545 |
25 |
605 |
70 |
450-550 | 570-700 |
TurboFP650 | 592 |
650 |
560 |
40 |
630 |
75 |
500-570 | 595-750 |
wtGFP | 474 |
509 |
450 |
50 |
525 |
50 |
370-470 | 490-700 |
X-Rhod-1 Indicator | 580 |
601 |
560 |
40 |
630 |
75 |
500-570 | 595-750 |
ZsGreen1 | 493 |
505 |
470 |
40 |
525 |
50 |
400-480 | 500-600 |
ZsYellow1 | 529 |
539 |
500 |
40 |
560 |
50 |
450-510 | 530-700 |
注:表格单位为nm