流式常用熒光染料

1、如何選擇流式細胞儀測定常用的熒光染料流式細胞儀測定常用的熒光染料有很多種,我們如何來選擇流式細胞儀測定的常用熒光染料呢？    首先要考慮，他們分子結構不同,激發(fā)光譜和發(fā)射光譜也不同,選擇熒光染料時，必須依據流式細胞儀所配備的激光光源的發(fā)射光波長做為重點考慮。發(fā)射光波長，如：氬離子氣體激光管,它的發(fā)射光波488nm；                氦氖離子氣體激光管發(fā)射光波長633nm；  

2、60;             488nm激光光源常用的熒光染料有FITC（異硫氰酸熒光素）；                PE（藻紅蛋白）               

3、; PI（碘化丙啶）                CY5（化青素）                preCP(葉綠素蛋白)              

4、;  ECD(藻紅蛋白-得克薩斯紅)等。 激發(fā)光和發(fā)射光波長，如下： 激發(fā)光波長（nm）發(fā)射光峰值（nm）FITC488525（綠）PE488575（橙紅）PI488630（橙紅）ECD488610（紅）CY5488675（深紅）PreCP488675（深紅）流式細胞儀中常用的熒光染料光學特征以及需要的激光器名    稱Ex波長(nm)Em波長(nm)   備    注激光器Hoechst 33342 355 465 DNA染色核酸標記

5、351nm紫外激光器Hoechst 33258365 465 DAPI372 456 Indo-1350 405 細胞內鈣離子標記FITC490 520 標記抗體488nm 氬 離 子 激 光 器R_PE480 578 PE_CY5480 670 PE_CY5.5480 695 PerCP490 675 Alexa Fluor 488 494 517 PE_

6、Texas Red 480 613 Rhodamine 123 500 540 線粒體標記Fluo-3506 526 細胞內鈣離子標記DiOC6(3)480 501 內質網標記YOYO-1490 510 DNA染色核酸標記Propidium Iodide530 615 Acridine Orange490 640 APC650 660 標記抗體633nm氦氖激光器Alexa Fluor 647 650 668 CY5

7、650 667 流式細胞儀交流論壇：熒光染料譜染料名稱Excitation(nm)Emission(nm)分子量備注信息Reactive and conjugated probesHydroxycoumarin325386331Succinimidyl esterAminocoumarin350445330Succinimidyl esterMethoxycoumarin360410317Succinimidyl esterCascade Blue(375);401423596HydrazidePacific Blue403455406MaleimidePacific Or

8、ange403551  Lucifer yellow425528  NBD466539294NBD-XR-Phycoerythrin (PE)480;565578240 k PE-Cy5 conjugates480;565;650670 aka Cychrome, R670, Tri-Color, Quantum RedPE-Cy7 conjugates480;565;743767  Red 613480;565613 PE-Texas RedPerCP490675 Peridinin chlo

9、rphyll proteinTruRed490,675695 PerCP-Cy5.5 conjugateFluorX494520587GE HealthcareFluorescein495519389FITC; pH sensitiveBODIPY-FL503512  TRITC547572444TRITCX-Rhodamine570576548XRITCLissamine Rhodamine B570590  Texas Red589615625Sulfonyl chlorideAllophycocyanin (APC)650660104 k

10、 APC-Cy7 conjugates650;755767 PharRedAlexa Fluor系列熒光染料Alexa Fluor 350343442410 Alexa Fluor 4054014211028 Alexa Fluor 430434540702 Alexa Fluor 488499519643QY 0.92Alexa Fluor 500503525700 Alexa Fluor 514517542714 Alexa Fluor 532530555724QY 0.61Alexa Fluor 54656157210

11、79QY 0.79Alexa Fluor 5555535681250QY 0.1Alexa Fluor 568579603792QY 0.69Alexa Fluor 594591618820QY 0.66Alexa Fluor 6106106291285 Alexa Fluor 6336326481200 Alexa Fluor 6476526681300QY 0.33Alexa Fluor 6606636911100 Alexa Fluor 6806807021150 Alexa Fluor 7006967191400 Alexa Fluor

12、 7507527761300 Alexa Fluor 7907828041750 Cy系列熒光染料Cy2489506714QY 0.12Cy3(512);550570;(615)767QY 0.15Cy3B558572;(620)658QY 0.67Cy3.5581594;(640)1102QY 0.15Cy5(625);650670792QY 0.28Cy5.56756941128QY 0.23Cy7743767818QY 0.28核酸相關探針染料Hoechst 33342343483616 AT-selective DAPI345455 AT-selectiv

13、e Hoechst 33258345478624AT-selective SYTOX Blue431480400DNA Chromomycin A3445575 CG-selective Mithramycin445575  YOYO#160;Ethidium Bromide493620394  Acridine Orange503530/640 DNA/RNASYTOX Green504523600 DNA TOTO-1, TO-PRO-1509533 Vital stain, TOTO: Cyanine

14、Dimer TO-PRO: Cyanine MonomerThiazole Orange510530  Propidium Iodide (PI)536617668.4 LDS 751543;590712;607472 DNA (543ex/712em), RNA (590ex/607em) 7-AAD546647 7-aminoactinomycin D, CG-selectiveSYTOX Orange547570500 DNA TOTO-3, TO-PRO-3642661  DRAQ5647681,697413usable ex

15、citation down to 488細胞功能探針I(yè)ndo-1361/330490/4051010AM ester. Low/High Ca+,Fluo-3506526855AM ester. pH > 6DCFH5055355292'7'Dichorodihydrofluorescein, oxidized formDHR505534346Dihydrorhodamine 123, oxidized form, light catalyzes oxidationSNARF548/579587/635 pH 6/9熒光蛋白   &

16、#160;QYBRPS Y66H360442     Y66F360508     EBFP380440 0.180.27 monomerEBFP2383448  20 monomerAzurite383447  15 monomerGFPuv385508     T-Sapphire399511 0.602625weak dimerCerulea

17、n433475 0.622736weak dimermCFP433475 0.401364monomerECFP434477 0.153  CyPet435477 0.511859weak dimerY66W436485     mKeima-Red440620 0.243 monomerTagCFP458480  29 dimerAmCyan1458489 0.7529 tetramermTFP1462492

18、60; 54 dimerS65A471504     Midoriishi Cyan472495 0.925 dimerWild Type GFP396,47550826k0.77   S65C479507     TurboGFP48250226 k0.5337 dimerTagGFP482505  34 monomerS65L484510    

19、; Emerald487509 0.68390.69weak dimer; (Invitrogen)S65T488511     EGFP48850726k 0.6034174weak dimerAzami Green492505 0.7441 monomerZsGreen1493505105k0.9140 tetramerTagYFP508524  47 monomerEYFP51452726k0.615160weak dimerTopaz514527 

20、 57 monomerVenus515528 0.575315weak dimermCitrine516529 0.765949monomerYPet517530 0.778049weak dimerTurboYFP52553826 k0.531.65 dimerZsYellow1529539 0.6513 tetramerKusabira Orange548559 0.6031 monomermOrange548562 0.69499monomermKO548559 0.6031122monomerTurboRFP55357426 k0.6762 dimertdTomato554581 0.699598tandem di