流式常用熒光染料

1、如何選擇流式細(xì)胞儀測定常用的熒光染料流式細(xì)胞儀測定常用的熒光染料有很多種,我們?nèi)绾蝸磉x擇流式細(xì)胞儀測定的常用熒光染料呢？ 首先要考慮，他們分子結(jié)構(gòu)不同,激發(fā)光譜和發(fā)射光譜也不同,選擇熒光染料時(shí)，必須依據(jù)流式細(xì)胞儀所配備的激光光源的發(fā)射光波長做為重點(diǎn)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；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） PI（碘化丙啶）CY5（化青素） preCP(葉綠素蛋白) 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）FITC

2、488525（綠）PE488575（橙紅）PI488630（橙紅）ECD488610（紅）CY5488675（深紅）PreCP488675（深紅）流式細(xì)胞儀中常用的熒光染料光學(xué)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 355465DNA染色核酸標(biāo)記351nm紫外激光器Hoechst 33258365465DAPI372456Indo-1350405細(xì)胞內(nèi)鈣離子標(biāo)記FITC490520標(biāo)記抗體488nm氬離子激光器R_PE480578PE_CY5480670PE_CY5.5480695PerCP490675Alexa Fluor 488 494

3、517PE_Texas Red 480613Rhodamine 123 500540線粒體標(biāo)記Fluo-3506526細(xì)胞內(nèi)鈣離子標(biāo)記DiOC6(3)480501內(nèi)質(zhì)網(wǎng)標(biāo)記YOYO-1490510DNA染色核酸標(biāo)記Propidium Iodide530615Acridine Orange490640APC650660標(biāo)記抗體633nm氦氖激光器Alexa Fluor 647 650668CY5650667流式細(xì)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 probesHydroxycoumari

4、n325386331Succinimidyl esterAminocoumarin350445330Succinimidyl esterMethoxycoumarin360410317Succinimidyl esterCascade Blue(375);401423596HydrazidePacific Blue403455406MaleimidePacific Orange403551Lucifer yellow425528NBD466539294NBD-XR-Phycoerythrin (PE)480;565578240 kPE-Cy5 conjugates480;565;650670a

5、ka Cychrome, R670, Tri-Color, Quantum RedPE-Cy7 conjugates480;565;743767Red 613480;565613PE-Texas RedPerCP490675Peridinin chlorphyll proteinTruRed490,675695PerCP-Cy5.5 conjugateFluorX494520587GE HealthcareFluorescein495519389FITC; pH sensitiveBODIPY-FL503512TRITC547572444TRITCX-Rhodamine570576548XRI

6、TCLissamine Rhodamine B570590Texas Red589615625Sulfonyl chlorideAllophycocyanin (APC)650660104 kAPC-Cy7 conjugates650;755767PharRedAlexa Fluor系列熒光染料Alexa Fluor 350343442410Alexa Fluor 4054014211028Alexa Fluor 430434540702Alexa Fluor 488499519643QY 0.92Alexa Fluor 500503525700Alexa Fluor 514517542714

7、Alexa Fluor 532530555724QY 0.61Alexa Fluor 5465615721079QY 0.79Alexa Fluor 5555535681250QY 0.1Alexa Fluor 568579603792QY 0.69Alexa Fluor 594591618820QY 0.66Alexa Fluor 6106106291285Alexa Fluor 6336326481200Alexa Fluor 6476526681300QY 0.33Alexa Fluor 6606636911100Alexa Fluor 6806807021150Alexa Fluor

8、7006967191400Alexa Fluor 7507527761300Alexa Fluor 7907828041750Cy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(guān)探針染料Hoechst 33342343483616 AT-selective DAPI345455AT-se

9、lective Hoechst 33258345478624AT-selective SYTOX Blue431480400DNA Chromomycin A3445575CG-selective Mithramycin445575YOYOthidium Bromide493620394 Acridine Orange503530/640DNA/RNASYTOX Green504523600 DNA TOTO-1, TO-PRO-1509533Vital stain, TOTO: Cyanine Dimer TO-PRO: Cyanine MonomerThiazol

10、e Orange510530Propidium Iodide (PI)536617668.4LDS 751543;590712;607472 DNA (543ex/712em), RNA (590ex/607em) 7-AAD5466477-aminoactinomycin D, CG-selectiveSYTOX Orange547570500 DNA TOTO-3, TO-PRO-3642661DRAQ5647681,697413usable excitation down to 488細(xì)胞功能探針I(yè)ndo-1361/330490/4051010AM ester. Low/High Ca+

11、,Fluo-3506526855AM ester. pH 6DCFH50553552927Dichorodihydrofluorescein, oxidized formDHR505534346Dihydrorhodamine 123, oxidized form, light catalyzes oxidationSNARF548/579587/635pH 6/9熒光蛋白QYBRPSY66H360442Y66F360508EBFP3804400.180.27monomerEBFP238344820monomerAzurite38344715monomerGFPuv385508T-Sapphi

12、re3995110.602625weak dimerCerulean4334750.622736weak dimermCFP4334750.401364monomerECFP4344770.153CyPet4354770.511859weak dimerY66W436485mKeima-Red4406200.243monomerTagCFP45848029dimerAmCyan14584890.7529tetramermTFP146249254dimerS65A471504Midoriishi Cyan4724950.925dimerWild Type GFP396,47550826k0.77

13、S65C479507TurboGFP48250226 k0.5337dimerTagGFP48250534monomerS65L484510Emerald4875090.68390.69weak dimer; (Invitrogen)S65T488511EGFP48850726k 0.6034174weak dimerAzami Green4925050.7441monomerZsGreen1493505105k0.9140tetramerTagYFP50852447monomerEYFP51452726k0.615160weak dimerTopaz51452757monomerVenus5

14、155280.575315weak dimermCitrine5165290.765949monomerYPet5175300.778049weak dimerTurboYFP52553826 k0.531.65dimerZsYellow15295390.6513tetramerKusabira Orange5485590.6031monomermOrange5485620.69499monomermKO5485590.6031122monomerTurboRFP55357426 k0.6762dimertdTomato5545810.699598tandem dimerTagRFP55558450monomerDsRed monomer55658628k0.13.516monomerDsRed2 (RFP)563582110k0.5524mStrawberry5745960.292615monomerTurboFP60257460226 k0.3526dimerAsRed2576592110k0.2113tetramermRFP158460730k0.25monomerJ-Red5846100.208.813dimermCherry5876100.221696monomerHcRed158861852k0.030