流式常用熒光染料概要

1、如何選擇流式細(xì)胞儀測(cè)定常用的熒光染料流式細(xì)胞儀測(cè)定常用的熒光染料有很多種，我們?nèi)绾蝸?lái)選擇流式細(xì)胞儀測(cè)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(zhǎng)做為重點(diǎn)考慮。發(fā)射光波長(zhǎng)，如：鼠離子氣體激光管，它的發(fā)射光波488nm氨窺離子氣體激光管發(fā)射光波長(zhǎng)633nm；488nm激光光源常用的熒光染料有FITC（異硫富酸熒光素）；PE（藻紅蛋白）PI（碘化丙咤）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(zhǎng)，如下:激發(fā)光波長(zhǎng)（nm發(fā)射光峰值（nm>FITC48852

2、5（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(zhǎng)(nm)Em波長(zhǎng)(nm)備注徼光器Hoechst33342355465DNA染色Hoechst33258365465351nm核酸標(biāo)記DAPI372456紫外激光器ndo-1350405細(xì)胞內(nèi)鈣離子標(biāo)記FITC490520R_PE480578PE_CY5480670PE_CY5.5480695標(biāo)記抗體488nmPerCP490675瀛AlexaFluor488494517離PE_TexasRed48

3、0613_子Rhodamine123500|540線粒體標(biāo)記激Fluo-3506|526細(xì)胞內(nèi)鈣離子標(biāo)記光DiOC6(3)480J內(nèi)質(zhì)網(wǎng)標(biāo)記崩YOYO-1490510PropidiumIodide530615DNA朱巴rnL_II核酸標(biāo)記AcridineOrange490640APC650660AlexaFluor647RsnRRR650668標(biāo)記抗體633nn氮鏡激光器CY5650667流式0田胞儀交流論壇：熒光染料譜染料名稱Excitation(nm)Emission(nm)分子量備注信息ReactiveandconjugatedprobesHydroxycoumarin32538633S

4、uccinimidylester1Aminocoumarin350445330SuccinimidylesterMethoxycoumarin360410317SuccinimidylesterCascadeBlue(375);401423596HydrazidePacificBlue403455406MaleimidePacificOrange403551Luciferyellow425528NBD466539294NBD-XR-Phycoerythrin(PE)480;565578240kPE-Cy5conjugates480;565;650670akaCychrome,R670,Tri-

5、Color,QuantumRedPE-Cy7conjugates480;565;743767Red613480;565613PE-TexasRedPerCP490675PeridininchlorphyllproteinTruRed490,675695PerCP-Cy5.5conjugateFluorX494520587GEHealthcareFluorescein495519389FITC;pHsensitiveBODIPY-FL503512TRITC547572444TRITCX-Rhodamine57057654XRITC8LissamineRhodamineB570590TexasRe

6、d589615625SulfonylchlorideAllophycocyanin(APC)650660104kAPC-Cy7conjugates650;755767PharRedAlexaFluor系列熒光染料AlexaFluor350343442410AlexaFluor4054014211028AlexaFluor430434540702AlexaFluor488499519643QY0.92AlexaFluor500503525700AlexaFluor514517542714AlexaFluor532530555724QY0.61AlexaFluor5465615721079QY0.

7、79AlexaFluor5555535681250QY0.1AlexaFluor568579603792QY0.69AlexaFluor594591618820QY0.66AlexaFluor6106106291285AlexaFluor6336326480012AlexaFluor6476526680013QY0.33AlexaFluor6606636910011AlexaFluor6806807025011AlexaFluor7006967190014AlexaFluor7507527760013AlexaFluor7907828045017Cy系列熒光染料Cy2489506471QY0.

8、12Cy3(512);550570;(615)776QY0.15Cy3B558572;(620)865QY0.67Cy3.5581594;(640)0211QY0.15Cy5(625);650670279QY0.28Cy5.56756942811QY0.23Cy7743767881QY0.28核酸相關(guān)探針染料Hoechst33342343483616AT-selectiveDAPI345455AT-selectiveHoechst33258345478624AT-selectiveSYTOXBlue431480400DNAChromomycinA3445575CG-selectiveMithr

9、amycin445575YOYOthidiumBromide493620394AcridineOrange503530/640DNA/RNASYTOXGreen504523600DNATOTO-1,TO-PRO-1509533Vitalstain,TOTO:CyanineDimerTO-PRO:CyanineMonomerThiazoleOrange510530PropidiumIodide(PI)536617668.4LDS751543;590712;607472DNA(543ex/712em),RNA(590ex/607em)7-AAD5466477-aminoa

10、ctinomycinD,CG-selectiveSYTOXOrange547570500DNATOTO-3,TO-PRO-3642661DRAQ5647681,697413usableexcitationdownto488細(xì)胞功能探針I(yè)ndo-1361/330490/4051010AMester.Low/HighCa+,Fluo-3506526855AMester.pH>6DCFH5055355292'7'Dichorodihydrofluorescein,oxidizedformDHR505534346Dihydrorhodamine123,oxidizedform,l

11、ightcatalyzesoxidationSNARF548/579587/635pH6/9熒光蛋白1JUJQYBRPSY66H360442Y66F360508EBFP3804400.180.27monomerEBFP238344820monomerAzurite38344715monomerGFPuv385508T-Sapphire3995110.602625weakdimerCerulean4750.622736weakdimermCFP4750.401364monomerECFP4770.153CyPet4770.511859weakdimerY66W485mKeima-Red16200

12、.243monomerTagCFP45848029dimerAmCyanl4584890.7529tetramermTFPI46249254dimerS65A471504MidoriishiCyan4724950.925dimerWildTypeGFP396,47550826k0.77S65C4791507TurboGFP48250226k0.5337dimerTagGFP48250534monomerS65L484510Emerald4875090.68390.69weakdimer;(Invitrogen)S65T488511EGFP48850726k0.6034174weakdimerA

13、zamiGreen4925050.7441monomerZsGreen1493505105k0.9140tetramerTagYFP50852447monomerEYFP514527260.56weakdimerk6110Topaz51452757monomerVenus5155280.575315weakdimermCitrine5165290.765949monomerYPet5175300.778049weakdimerTurboYFP52553826k0.531.65dimerZsYellowl5295390.6513tetramerKusabiraOrange5485590.6031

14、monomermOrange5485620.69499monomermKO5485590.6031122monomerTurboRFP55357426k0.6762dimertdTomato5545810.699598tandemdimerTagRFP55558450monomerDsRedmonomer55658628k0.13.516monomerDsRed2("RFP")563582110k0.5524mStrawberry5745960.292615monomerTurboFP602574602260.2dimerk356AsRed2576592110k0.2113tetramermRFPI58460730k0.25monomerJ-Red5846100.208.813dimermCherry5876100.221696monomerHcRedl58861852k0.030.6dim