流式常用荧光染料概要.docx

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633nm488nm激光光源常用的荧光染料有FITC （异硫氧酸荧光素）；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（nmFITC488525 （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:x波长（nm）激光器Hoechst 33342Hoechst 33258DAPIndo-1F|T CPE_CY5480PE_CY5.5480PerCP490Alexa Fluor 488494PE_Texas Red480Rhodamine 123500Fluo-3506DiOC6(3)480PE8090Em波长(nm)355365372350490540 ■线粒体标记■ 510细胞内钙离子标记内质网标记351nm紫外激光器488nmYOYO-1ropidium IodideAcridine OrangeAPCAlexa Fluor 647CY5530615DNA染色633nm氦氖激光器流式掘田胞仪交流论坛. ml荧光染料谱染料名称Excitation(nm)Emission(nm)分子量备注信息Reactive and conjugated probesHydroxycoumarin32538633Succinimidyl esterSuccinimidyl esterSuccinimidyl esterCascade BlueHydrazidePacific BluePacific OrangeLucifer yellowR-Phycoerythrin (PE)480;565;6PE-Cy5 conjugates50Red480;565;7PE-Cy7 conjugates43Red 613PE-Texas RedPerCPPeridinin chlorphyll proteinPerCP-Cy5.5 conjugateTruRedGE HealthcareFluorXFITC; pH sensitiveFluoresceinBODIPY-FLMethoxycoumarinX-Rhodamine480;565480;565490,675aka Cychrome, R670, Tri-Color, QuantumLissamineRhodamine BTexas RedAllophycocyanin(APC)APC-Cy7 conjugatesAlexa Fluor系列荧光染料Alexa Fluor 350Alexa Fluor 405Alexa Fluor 430Alexa Fluor 488Alexa Fluor 500Alexa Fluor 514Alexa Fluor 532Alexa Fluor 546Alexa Fluor 555Alexa Fluor 568Alexa Fluor 594Alexa Fluor 610QY 0.1QY 0.79QY 0.6985Alexa Fluor 6336326481200Alexa Fluor 6476526681300QY 0.33Alexa Fluor 6606636911100Alexa Fluor 6806807021150Alexa Fluor 7006967191400Alexa Fluor 7507527761300Alexa Fluor 7907828041750Cy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 33342AT-selectiveAT-selectiveHoechst 33258AT-selectiveSYTOX BlueChromomycin A3Mithramycin12CG-selectiveAcridine OrangeSYTOX GreenTOTO-1, TO-PRO-1TO-PRO: Cyanine MonomerThiazole OrangePropidium Iodide (PI)Ethidium BromideDNA/RNAVital stain, TOTO: Cyanine Dimer7-aminoactinomycin D, CG-selective细胞功能探针SYTOX OrangeTOTO-3, TO-PRO-3Indo-1361/330490/4051010AM ester. Low/High Ca++,Fluo-3506526855AM ester. pH > 6DCFH5055355292'7'Dichorodihydrofluorescein, oxidized formDHR505534346Dihydrorhodamine 123, oxidized form, light catalyzes oxidationSNARF548/579587/635pH 6/9荧光蛋白BRPSY66H36508Y66FEBFP380 440EBFP2383 448Azurite383447GFPuv385508T-Sapphire399511CeruleanmCFP475475ECFP477CyPet477Y66W48500.monomer18271 1I~20monomer□monomer1 >10.22weak dimer60650.23I"weak djm,er62761 0.16monome，4034r0.3150.15weak dimer518mKeima-RedTagCFPAmCyanltetramermTFPIMidoriishi CyanWild Type GFP(Invitrogen)weak dimermonomertetramermonomerweak dimerEmeraldAzami GreenZsGreen1396,475weak dimer;k6110Topaz5145275monomerVenus5155280.5755 1weak dimermCitrine5165290.7654monomerYPet517530。