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离子成像
在有关神经元方面的研究中,观察基因活性或细胞运动等对于了解细胞的离子浓度具有重要意义。钠、钙、氯或镁离子对很多不同的细胞活动都具有较大影响。一般情况下,借助荧光标记的螯合剂可将离子困住,螯合剂在结合相应离子后会改变离子的光谱特性。例如,利用该原理的钙指示剂fura-2、indo-1、fluo-3、fluo-4和Calcium-Green 等。
对于钠离子的检测,通常使用 SBFI (sodium-bindingbenzofurzanisophthalate) 或Sodium Green。PBFI(potassium-binding benzofurzanisophthalate) 可以检测钾离子。
有趣的是,还存在以蛋白质为主的钙指示剂,其中一种是基于水母化学发光蛋白—水母素。水母素、发光体腔肠素、分子氧和 Ca2+的相互作用会释放蓝光,这是在荧光蛋白质的发现过程中非常著名的机制。
荧光染料及其激发和发射波长峰值
上文提到的所有染料在下表中均有统计。此外,还涵盖了其他荧光染料及其激发和发射波长峰值。
荧光染料示例 |
激发光 |
Indo-1, Ca saturated |
331 nm |
Indo-1 Ca2+ |
346 nm |
Cascade Blue BSA pH 7.0 |
401 nm |
Cascade Blue |
398 nm |
LysoTracker Blue |
373 nm |
Alexa 405 |
401 nm |
LysoSensor Blue pH 5.0 |
374 nm |
LysoSensor Blue |
374 nm |
DyLight 405 |
399 nm |
DyLight 350 |
332 nm |
BFP (Blue Fluorescent Protein) |
380 nm |
Alexa 350 |
343 nm |
7-Amino-4-methylcoumarin pH 7.0 |
346 nm |
Amino Coumarin |
345 nm |
AMCA conjugate |
347 nm |
Coumarin |
360 nm |
7-Hydroxy-4-methylcoumarin |
360 nm |
7-Hydroxy-4-methylcoumarin pH 9.0 |
361 nm |
6,8-Difluoro-7-hydroxy-4-methylcoumarin pH 9.0 |
358 nm |
Hoechst 33342 |
352 nm |
Pacific Blue |
404 nm |
Hoechst 33258 |
352 nm |
Hoechst 33258-DNA |
352 nm |
Pacific Blue antibody conjugate pH 8.0 |
404 nm |
PO-PRO-1 |
434 nm |
PO-PRO-1-DNA |
435 nm |
POPO-1 |
433 nm |
POPO-1-DNA |
433 nm |
DAPI-DNA |
359 nm |
DAPI |
358 nm |
Marina Blue |
362 nm |
SYTOX Blue-DNA |
445 nm |
CFP (Cyan Fluorescent Protein) |
434 nm |
eCFP (Enhanced Cyan Fluorescent Protein) |
437 nm |
1-Anilinonaphthalene-8-sulfonic acid (1,8-ANS) |
375 nm |
Indo-1, Ca free |
346 nm |
1,8-ANS (1-Anilinonaphthalene-8-sulfonic acid) |
375 nm |
BO-PRO-1-DNA |
462 nm |
BOPRO-1 |
462 nm |
BOBO-1-DNA |
461 nm |
SYTO 45-DNA |
451 nm |
evoglow-Pp1 |
448 nm |
evoglow-Bs1 |
448 nm |
evoglow-Bs2 |
448 nm |
Auramine O |
431 nm |
DiO |
487 nm |
LysoSensor Green pH 5.0 |
447 nm |
Cy 2 |
489 nm |
LysoSensor Green |
447 nm |
Fura-2, high Ca |
336 nm |
Fura-2 Ca2+sup> |
336 nm |
SYTO 13-DNA |
488 nm |
YO-PRO-1-DNA |
491 nm |
YOYO-1-DNA |
491 nm |
eGFP (Enhanced Green Fluorescent Protein) |
488 nm |
LysoTracker Green |
503 nm |
GFP (S65T) |
489 nm |
BODIPY FL, MeOH |
502 nm |
Sapphire |
396 nm |
BODIPY FL conjugate |
503 nm |
MitoTracker Green |
490 nm |
MitoTracker Green FM, MeOH |
490 nm |
Fluorescein 0.1 M NaOH |
493 nm |
Calcein pH 9.0 |
494 nm |
Fluorescein pH 9.0 |
490 nm |
Calcein |
493 nm |
Fura-2, no Ca |
367 nm |
Fluo-4 |
494 nm |
FDA |
495 nm |
DTAF |
495 nm |
Fluorescein |
495 nm |
Fluorescein antibody conjugate pH 8.0 |
493 nm |
CFDA |
495 nm |
FITC |
495 nm |
Alexa Fluor 488 hydrazide-water |
493 nm |
DyLight 488 |
493 nm |
5-FAM pH 9.0 |
492 nm |
FITC antibody conjugate pH 8.0 |
495 nm |
Alexa 488 |
493 nm |
Rhodamine 110 |
497 nm |
Rhodamine 110 pH 7.0 |
497 nm |
Acridine Orange |
431 nm |
Alexa Fluor 488 antibody conjugate pH 8.0 |
499 nm |
BCECF pH 5.5 |
485 nm |
PicoGreendsDNA quantitation reagent |
502 nm |
SYBR Green I |
498 nm |
Rhodaminen Green pH 7.0 |
497 nm |
CyQUANT GR-DNA |
502 nm |
NeuroTrace 500/525, green fluorescent Nissl stain-RNA |
497 nm |
DansylCadaverine |
335 nm |
Rhodol Green antibody conjugate pH 8.0 |
499 nm |
Fluoro-Emerald |
495 nm |
Nissl |
497 nm |
Fluorescein dextran pH 8.0 |
501 nm |
Rhodamine Green |
497 nm |
5-(and-6)-Carboxy-2', 7'-dichlorofluorescein pH 9.0 |
504 nm |
DansylCadaverine, MeOH |
335 nm |
eYFP (Enhanced Yellow Fluorescent Protein) |
514 nm |
Oregon Green 488 |
498 nm |
Oregon Green 488 antibody conjugate pH 8.0 |
498 nm |
Fluo-3 |
506 nm |
BCECF pH 9.0 |
501 nm |
SBFI-Na+ |
336 nm |
Fluo-3 Ca2+ |
506 nm |
Rhodamine 123, MeOH |
507 nm |
FlAsH |
509 nm |
Calcium Green-1 Ca2+ |
506 nm |
Magnesium Green |
507 nm |
DM-NERF pH 4.0 |
493 nm |
Calcium Green |
506 nm |
Citrine |
515 nm |
LysoSensor Yellow pH 9.0 |
335 nm |
TO-PRO-1-DNA |
515 nm |
Magnesium Green Mg2+ |
507 nm |
Sodium Green Na+ |
507 nm |
TOTO-1-DNA |
514 nm |
Oregon Green 514 |
512 nm |
Oregon Green 514 antibody conjugate pH 8.0 |
513 nm |
NBD-X |
466 nm |
DM-NERF pH 7.0 |
509 nm |
NBD-X, MeOH |
467 nm |
CI-NERF pH 6.0 |
513 nm |
Alexa 430 |
431 nm |
Alexa Fluor 430 antibody conjugate pH 7.2 |
431 nm |
CI-NERF pH 2.5 |
504 nm |
Lucifer Yellow, CH |
428 nm |
LysoSensor Yellow pH 3.0 |
389 nm |
6-TET, SE pH 9.0 |
521 nm |
Eosin antibody conjugate pH 8.0 |
525 nm |
Eosin |
524 nm |
6-Carboxyrhodamine 6G pH 7.0 |
526 nm |
6-Carboxyrhodamine 6G, hydrochloride |
525 nm |
Bodipy R6G SE |
528 nm |
BODIPY R6G, MeOH |
528 nm |
6 JOE |
520 nm |
Cascade Yellow antibody conjugate pH 8.0 |
399 nm |
Cascade Yellow |
399 nm |
mBanana |
540 nm |
Alexa Fluor 532 antibody conjugate pH 7.2 |
528 nm |
Alexa 532 |
528 nm |
Erythrosin-5-isothiocyanate pH 9.0 |
533 nm |
6-HEX, SE pH 9.0 |
534 nm |
mOrange |
548 nm |
mHoneydew |
478 nm |
Cy 3 |
549 nm |
Rhodamine B |
543 nm |
DiI |
551 nm |
5-TAMRA-MeOH |
543 nm |
Alexa 555 |
553 nm |
Alexa Fluor 555 antibody conjugate pH 7.2 |
553 nm |
DyLight 549 |
555 nm |
BODIPY TMR-X, SE |
544 nm |
BODIPY TMR-X, MeOH |
544 nm |
PO-PRO-3-DNA |
539 nm |
PO-PRO-3 |
539 nm |
Rhodamine |
551 nm |
Bodipy TMR-X conjugate |
544 nm |
POPO-3 |
533 nm |
Alexa 546 |
562 nm |
BODIPY TMR-X antibody conjugate pH 7.2 |
544 nm |
Calcium Orange Ca2+ |
549 nm |
TRITC |
550 nm |
Calcium Orange |
549 nm |
Rhodaminephalloidin pH 7.0 |
558 nm |
MitoTracker Orange |
551 nm |
MitoTracker Orange, MeOH |
551 nm |
Phycoerythrin |
565 nm |
Magnesium Orange |
550 nm |
R-Phycoerythrin pH 7.5 |
565 nm |
5-TAMRA pH 7.0 |
553 nm |
5-TAMRA |
549 nm |
Rhod-2 |
552 nm |
FM 1-43 |
472 nm |
Rhod-2 Ca2+ |
553 nm |
Tetramethylrhodamine antibody conjugate pH 8.0 |
552 nm |
FM 1-43 lipid |
473 nm |
LOLO-1-DNA |
568 nm |
dTomato |
554 nm |
DsRed |
563 nm |
Dapoxyl (2-aminoethyl) sulfonamide |
372 nm |
Tetramethylrhodamine dextran pH 7.0 |
555 nm |
Fluor-Ruby |
554 nm |
Resorufin |
571 nm |
Resorufin pH 9.0 |
571 nm |
mTangerine |
568 nm |
LysoTracker Red |
578 nm |
Lissaminerhodamine |
572 nm |
Cy 3.5 |
578 nm |
Rhodamine Red-X antibody conjugate pH 8.0 |
573 nm |
Sulforhodamine 101, EtOH |
578 nm |
JC-1 pH 8.2 |
593 nm |
JC-1 |
592 nm |
mStrawberry |
575 nm |
MitoTracker Red |
578 nm |
MitoTracker Red, MeOH |
578 nm |
X-Rhod-1 Ca2+ |
580 nm |
Alexa Fluor 568 antibody conjugate pH 7.2 |
579 nm |
Alexa 568 |
576 nm |
5-ROX pH 7.0 |
578 nm |
5-ROX (5-Carboxy-X-rhodamine, triethylammonium salt) |
578 nm |
BO-PRO-3-DNA |
574 nm |
BOPRO-3 |
574 nm |
BOBO-3-DNA |
570 nm |
Ethidium Bromide |
524 nm |
ReAsH |
597 nm |
Calcium Crimson |
589 nm |
Calcium Crimson Ca2+ |
590 nm |
mRFP |
585 nm |
mCherry |
587 nm |
Texas Red-X antibody conjugate pH 7.2 |
596 nm |
HcRed |
590 nm |
DyLight 594 |
592 nm |
Ethidium homodimer-1-DNA |
528 nm |
Ethidiumhomodimer |
528 nm |
Propidium Iodide |
538 nm |
SYPRO Ruby |
467 nm |
Propidium Iodide-DNA |
538 nm |
Alexa 594 |
590 nm |
BODIPY TR-X, SE |
588 nm |
BODIPY TR-X, MeOH |
588 nm |
BODIPY TR-X phallacidin pH 7.0 |
590 nm |
Alexa Fluor 610 R-phycoerythrin streptavidin pH 7.2 |
567 nm |
YO-PRO-3-DNA |
613 nm |
Di-8 ANEPPS |
469 nm |
Di-8-ANEPPS-lipid |
469 nm |
YOYO-3-DNA |
612 nm |
Nile Red-lipid |
553 nm |
Nile Red |
559 nm |
DyLight 633 |
624 nm |
mPlum |
587 nm |
TO-PRO-3-DNA |
642 nm |
DDAO pH 9.0 |
648 nm |
Fura Red, high Ca |
434 nm |
Allophycocyanin pH 7.5 |
651 nm |
APC (allophycocyanin) |
650 nm |
Nile Blue, EtOH |
631 nm |
TOTO-3-DNA |
642 nm |
Cy 5 |
646 nm |
BODIPY 650/665-X, MeOH |
646 nm |
Alexa Fluor 647 R-phycoerythrin streptavidin pH 7.2 |
569 nm |
DyLight 649 |
652 nm |
Alexa Fluor 647 antibody conjugate pH 7.2 |
653 nm |
Alexa 647 |
653 nm |
Fura Red Ca2+ |
435 nm |
Atto 647 |
644 nm |
Fura Red, low Ca |
472 nm |
Carboxynaphthofluorescein pH 10.0 |
600 nm |
Alexa 660 |
664 nm |
Alexa Fluor 660 antibody conjugate pH 7.2 |
663 nm |
Cy 5.5 |
673 nm |
Alexa Fluor 680 antibody conjugate pH 7.2 |
679 nm |
Alexa 680 |
679 nm |
DyLight 680 |
678 nm |
Alexa Fluor 700 antibody conjugate pH 7.2 |
696 nm |
Alexa 700 |
696 nm |
FM 4-64, 2% CHAPS |
506 nm |
FM 4-64 |
508 nm |
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