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；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）PI（碘化丙啶）CY5（化青素）preCP（叶绿素蛋白）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Hoechst33342355465DNA染色核酸标记351nm紫外激光器Hoechst33258365465DAPI372456IndoT350405细胞内钙离子标记FITC490520标记抗体488nm氩离子激光器R_PE480578PE_CY5480670PE_CY5.5480695PerCP490675AlexaFluor488494517PE_TexasRed480■613Rhodamine123500540线粒体标记Fluo-3506526细胞内钙离子标记DiOC6(3)480■501内质网标记YOYO-1490■510DNA染色核酸标记PropidiumIodide530615AcridineOrange490■640APC650660标记抗体633nm氦氖激光器AlexaFluor647650668CY5650667流式•细胞仪交流论坛：http://bbs.bbioo.com/forum-181-1・html荧光染料谱染料名称Excitation(nm)Emission(nm)分子量备注信息ReactiveandconjugatedprobesHydroxycoumarin325386331SuccinimidylesterAminocoumarin350445330SuccinimidylesterMethoxycoumarin360410317SuccinimidylesterCascadeBlue(375);401423596HydrazidePacificBlue403455406MaleimidePacificOrange403551Luciferyellow425528NBD466539294NBD-XR-Phycoerythrin(PE)480;565578240kPE-Cy5conjugates480;565;650670akaCychrome,R670,Tri-Color,QuantumRedPE-Cy7conjugates480;565;743767Red613480;565613PE-TexasRedPerCP490675PeridininchlorphyllproteinTruRed490,675695PerCP-Cy5.5conjugateFluorX494520587GEHealthcareFluorescein495519389FITC;pHsensitiveBODIPY-FL503512TRITC547572444TRITCX-Rhodamine570576548XRITCLissamineRhodamineB570590TexasRed58961562Sulfonylchloride5Allophycocyanin(APC)650660104kAPC-Cy7conjugates650;755767PharRedAlexaFluor系列荧光染料AlexaFluor35034344241AlexaFluor4054014211028AlexaFluor43043454070AlexaFluor48849951964QY0.92AlexaFluor50050352570AlexaFluor51451754271AlexaFluor53253055572QY0.61AlexaFluor5465615721079QY0.79AlexaFluor5555535681250QY0.1AlexaFluor56857960379QY0.69AlexaFluor59459161882QY0.66AlexaFluor6106106291285AlexaFluor6336326481200343Hoechst33258345478462AT-selectiveSYTOXBlue43148000~4DNAChromomycinA3445575CG-selectiveMithramycin445575YOYO-14915097112EthidiumBromide493620439AcridineOrange503530/640DNA/RNASYTOXGreen50452300~6DNATOTO-1,TO-PRO-1509533Vitalstain,TOTO:CyanineDimerTO-PRO:CyanineMonomerThiazoleOrange510530PropidiumIodide(PI)5366178.466LDS751543;590712;607247DNA(543ex/712em),RNA(590ex/607em)7-AAD5466477-aminoactinomycinD,CG-selectiveSYTOXOrange54757000~5DNATOTO-3,TO-PRO-3642661DRAQ5647681,697341usableexcitationdownto488细胞功能探针Indo-1361/330490/4051010AMester.Low/HighCa++,Fluo-350652685AMester.pH>6550553552DCFH92'7'Dichorodihydrofluorescein,oxidizedformDHR505534346Dihydrorhodamine123,oxidizedform,lightcatalyzesoxidationSNARF548/579587/635pH6/9荧光蛋白QYBRPSY66H360442Y66F360508EBFP380440180.270.monomerEBFP238344802monomerAzurite38344751monomerGFPuv3855083995110.22T-Sapphire6065weakdimerCerulean433475620.7263weakdimermCFP433475400.3146monomerECFP434477150.3CyPet435477510.8195weakdimerY66W436485mKeima-Red440620240.3monomerTagCFP4584802dimer9AmCyanl4584890.752tetramermTFPI4624925dimerS65A471504MidoriishiCyan4724950.2dimerWildTypeGFP396,475508260.S65C479507TurboGFP482502260.533dimerTagGFP4825053monomerS65L484510Emerald4875090.6830.69weakdimer;(Invitrogen)S65T488511EGFP488507260.603741weakdimerAzamiGreen4925050.744monomerZsGreen1493505105k0.914tetramerTagYFP5085244monomerEYFP514527260.6156weakdimerTopaz5145275monomerVenus515528570.3551weakdimermCitrine516529760.9594monomerYPet517530770.0894weakdimerTurboYFP525538k26530.651.dimerZsYellowl529539650.31tetramer5485590.3KusabiraOrange601monomermOrange548562690.949monomermKO548559600.13221monomerTurboRFP553574k26670.26dimer5545810.99tdTomato6958tandemdimerTagRFP55558405monomer556586~20.3.1DsRedmonomer8k156monomerDsRed2("RFP")56358210k~1550.425745960.21mStrawberry2965monomerTurboFP602574602k26350.62dimerAsRed257659210k~1210.31tetramermRFPI5846070k~3250.monomerJ-Red584610200.88.31dimermCherry587610220.6169monomerHcRedl5886182k~5030.60.dimerKatusha58863532dimermKate(TagFP635)58863551monomerTurboFP635588635k26340.22dimermPlum590649100.14.35monomer;mRaspberry598625150.31fasterphotobleachthanmPlum其他探针Monochlorobimane380461622GlutathioneprobeCalcein496517362pH>5
