流式常用荧光染料

1、如何选择流式细胞仪测定常用的荧光染料流式细胞仪测定常用的荧光染料有很多种,我们如何来选择流式细胞仪测定的常用荧光染料呢？    首先要考虑，他们分子结构不同,激发光谱和发射光谱也不同,选择荧光染料时，必须依据流式细胞仪所配备的激光光源的发射光波长做为重点考虑。发射光波长，如：氩离子气体激光管,它的发射光波488nm；                氦氖离子气体激光管发射光波长633nm；  

2、60;             488nm激光光源常用的荧光染料有FITC（异硫氰酸荧光素）；                PE（藻红蛋白）               

3、; PI（碘化丙啶）                CY5（化青素）                preCP(叶绿素蛋白)              

4、;  ECD(藻红蛋白-得克萨斯红)等。 激发光和发射光波长，如下： 激发光波长（nm）发射光峰值（nm）FITC488525（绿）PE488575（橙红）PI488630（橙红）ECD488610（红）CY5488675（深红）PreCP488675（深红）流式细胞仪中常用的荧光染料光学特征以及需要的激光器名    称Ex波长(nm)Em波长(nm)   备    注激光器Hoechst 33342 355 465 DNA染色核酸标记

5、351nm紫外激光器Hoechst 33258365 465 DAPI372 456 Indo-1350 405 细胞内钙离子标记FITC490 520 标记抗体488nm 氩 离 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 线粒体标记Fluo-3506 526 细胞内钙离子标记DiOC6(3)480 501 内质网标记YOYO-1490 510 DNA染色核酸标记Propidium Iodide530 615 Acridine Orange490 640 APC650 660 标记抗体633nm氦氖激光器Alexa Fluor 647 650 668 CY5

7、650 667 流式细胞仪交流论坛：荧光染料谱染料名称Excitation(nm)Emission(nm)分子量备注信息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系列荧光染料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系列荧光染料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核酸相关探针染料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细胞功能探针Indo-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荧光蛋白   &

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