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：鼠离子气体激光管，它的发射光波488nm氨窥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 （异硫富酸荧光素）；PE（藻红蛋白）PI（碘化丙咤）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>FITC4

2、88525 （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 33342355465DNA染色核酸标记Hoechst 33258365465351nmDAPI372456紫外激光器ndo-1350405细胞内钙离子标记FITC490520R_PE480578PE_CY5480I1670PE_CY5.5480695标记抗体PerCP490675488nmAlexa Fluor 488494

3、517瀛离子PE_Texas Red480613Rhodamine 123500540线粒体标记激Fluo-3506526细胞内钙离子标记光器DiOC6(3)480501内质网标记YOYO-1490510DNA染色核酸标记Propidium Iodide530615Acridine Orange490640APC650I660633nm氮镜激光器Alexa Fluor 647650668标记抗体CY5650667流式借田胞仪交流论坛：http:荧光染料谱染料名称Excitation(nm)Emission(nm)分子量备注信息Reactive and conjugated probesHydr

4、oxycoumarin325386331Succinimidyl esterAminocoumarin350445330Succinimidyl esterMethoxycoumarin360410317Succinimidyl esterCascade Blue(375);401423596HydrazidePacific Blue403455406MaleimidePacific Orange403551Lucifer yellow425528NBD466539294NBD-XR-Phycoerythrin (PE)480;565578240 kPE-Cy5 conjugates480;5

5、65;650670aka Cychrome, R670, Tri-Color, QuantumRedPE-Cy7 conjugates480;565;743767Red 613480;565613PE-Texas RedPerCP490675Peridinin chlorphyll proteinTruRed490,675695PerCP-Cy5.5 conjugateFluorX494520587GE HealthcareFluorescein495519389FITC; pH sensitiveBODIPY-FL503512TRITC547572444TRITCX-Rhodamine570

6、576548XRITCLissamineRhodamine B570590Texas Red589615625Sulfonyl chlorideAllophycocyanin(APC)650660104 kAPC-Cy7 conjugates650;755767PharRedAlexa Fluor系列荧光染料Alexa Fluor 350343410Alexa Fluor 4054014211028Alexa Fluor 430434540702Alexa Fluor 488499519643QY 0.92Alexa Fluor 500503525700Alexa Fluor 51451754

7、2714Alexa Fluor 532530555724QY 0.61Alexa Fluor 54656157210QY 0.7979Alexa Fluor 5555535681250QY 0.1Alexa Fluor 568579603792QY 0.69Alexa Fluor 594591618820QY 0.66Alexa Fluor 6106106291285Alexa Fluor 6336326481200Alexa Fluor 6476526681300QY 0.33Alexa Fluor 6606636911100Alexa Fluor 6806807021150Alexa Fl

8、uor 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 33342343483616AT-selectiveDAPI345455AT-

9、selectiveHoechst 33258345478624AT-selectiveSYTOX Blue1480400DNAChromomycin A3445575CG-selectiveMithramycin445575YOYOthidium Bromide493620394Acridine Orange503530/640DNA/RNASYTOX Green504523600DNATOTO-1, TO-PRO-1509533Vital stain, TOTO: Cyanine DimerTO-PRO: Cyanine MonomerThiazole Orange

10、510530Propidium Iodide (PI)536617668.4LDS 751543;590712;607472DNA (543ex/712em), RNA (590ex/607em)7-AAD5466477-aminoactinomycin D, CG-selectiveSYTOX Orange547570500DNArOTO-3, TO-PRO-3642661DRAQ5647681,697413usable excitation down to 488细胞功能探针Indo-1361/330490/4051010AM ester. Low/High Ca+,Fluo-350652

11、6855AM ester. pH > 6DCFH5055355292'7'Dichorodihydrofluorescein, oxidized formDHR505534346Dihydrorhodamine 123, oxidized form, light catalyzes oxidationSNARF548/579587/635pH 6/9荧光蛋白QYBRPS丫66H360442Y66F360508EBFP3804400.180.27monomerEBFP238344820monomerAzurite38344715monomerGFPuv385508T-Sap

12、phire3995110.602625weak dimerCerulean4750.622736weak dimermCFP4750.401364monomerECFP4770.153CyPet4770.511859weak dimerY66W485mKeima-Red4406200.243monomerTagCFP45848029dimerAmCyanl4584890.7529tetramermTFPI46249254dimerS65A471504Midoriishi Cyan4724950.925dimerWild Type GFP396,47550826k0.77S65C479507Tu

13、rboGFP48250226k0.5337dimerTagGFP48250534monomerS65L484510Emerald4875090.68390.69weak dimer;(Invitrogen)S65T488511EGFP48850726k0.6034174weak dimerAzami Green4925050.7441monomerZsGreenl493505105k0.9140tetramerTagYFP50852447monomerEYFP51452726k0.615160weak dimerTopaz51452757monomerVenus5155280.575315we

14、ak dimermCitrine5165290.765949monomerYPet5175300.778049weak dimerTurboYFP52553826k0.531.65dimerZsYellow15295390.6513tetramerKusabira Orange5485590.6031monomermOrange5485620.69499monomermKO5485590.6031122monomerTurboRFP55357426k0.6762dimertdTomato5545810.699598tandem dimerTagRFP55558450monomerDsRed monomer55658628k0.13.516monomerDsRed2 ("RFP")563582110k0.5524mStrawberry5745960.292615monomerTurboFP60257460226k0.3526dimerAsRed2576592110k0.2113tetramermRFP158460730k0.25monomerJ-Red5846100.208.813dimermCherry5876100.221696monomerHcRed158861852k0.030