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：氯离子气体激光管，它的发射光波488nm氮氤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（异硫割酸荧光素）；PE（藻红蛋白）PI（碘化丙嚏）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

2、（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PEAlexa Fluor 488ETexas RedRhodamine 123Fluo-3Alexa Fluor 6476548048050050649050细胞内钙离子695510615660668内质网标记DNA 染色标记抗体流式乏田胞仪交流备 注激光器Ex波长(nm)m波长(nm)Hoechst 33342355465DNA染色Hoechst 33258465核酸标记351nmAPI372456

3、紫外激光器ndo-1350标记ITC490名 称论坛：http:/ and conjugated probesHydroxycoumarin325386331Succinimidyl esterAminocoumarin350445330Succinimidyl esterMethoxycoumarin360410317Succinimidyl esterCascade Blue(375);401423596HydrazidePacific Blue403455406MaleimidePacific Orange403551Lucifer yellow425528NBD466539294NBD

4、-XR-Phycoerythrin (PE)480;565578240 kPE-Cy5 conjugates480;565;650670aka Cychrome, R670, Tri-Color, QuantumRedPE-Cy7 conjugates480;565;743767Red 613480;565613PE-Texas RedPerCP490675Peridinin chlorphyll proteinTruRed490,675695PerCP-Cy5.5 conjugateFluorX494520587GE HealthcareFluorescein495519389FITC; p

5、H sensitiveBODIPY-FL503512TRITC547572444TRITCX-Rhodamine570576548XRITCLissamineRhodamine B570590Texas Red589615625Sulfonyl chlorideAllophycocyanin(APC)650660104 kAPC-Cy7 conjugates650;755767PharRedAlexa Fluor系列荧光染料Alexa Fluor 350343410Alexa Fluor 4054014211028Alexa Fluor 430434540702Alexa Fluor 4884

6、99519643QY 0.92Alexa Fluor 500503525700Alexa Fluor 514517542714Alexa 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 0.3

7、3Alexa 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

8、 0.28核酸相关探针染料Hoechst 33342343483616AT-selectiveDAPI345455AT-selectiveHoechst 33258345478624AT-selectiveSYTOX Blue431480400DNAChromomycin A3445575CG-selectiveMithramycin445575YOYOthidium Bromide493620394Acridine Orange503530/640DNA/RNASYTOX Green504523600DNATOTO-1, TO-PRO-1509533Vital st

9、ain, TOTO: Cyanine DimerTO-PRO: Cyanine MonomerThiazole Orange510530Propidium 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

10、功能探针Indo-1361/330490/4051010AM ester. Low/High Ca+,Fluo-3506526855AM ester. pH 6DCFH50553552927Dichorodihydrofluorescein, oxidized formDHR505534346Dihydrorhodamine 123, oxidized form, light catalyzesoxidationSNARF548/579587/635pH 6/9荧光蛋白QYBRPSY66H360442Y66F360508EBFP3804400.180.27monomerEBFP23834482

11、0monomerAzurite38344715monomerGFPuv385508T-Sapphire3995110.602625weak dimerCerulean4750.622736weak dimermCFPECFPCyPetY66WmKeima-RedTagCFP458AmCyan1458mTFP1462S65A471Midoriishi Cyan472Wild Type GFP396,4750.475monomer400.477150.477weak dimer51485620monomer24480dimer489tetramer75492dimer5040.495dimer26

12、0.508k77479507S65C26482502TurboGFPdimer53482505TagGFPmonomer484510S65Lweak dimer;487509Emerald6869488511S65T26488507EGFPweak dimer6074(Invitrogen)Azami Green4925050.7441monomerZsGreenl493505105k0.9140tetramerTagYFPEYFP50851452452726k0.61475160monomerweak dimerTopaz51452757monomerVenus5155280.575315w

13、eak dimermCitrine5165290.765949monomerYPet5175300.778049weak dimerTurboYFP52553826k0.531.65dimerZsYellow15295390.6513tetramerKusabira Orange5485590.6031monomermOrange5485620.69499 monomermKO5485590.6031122monomerTurboRFP55357426k0.6762dimertdTomato5545810.699598tandem dimerTagRFP55558450monomerDsRed monomer55658628k0.13.516monomerDsRed2 (RFP)563582110k0.5524mStrawberry5745960.292615monomerTurboFP60257460226k0.3526dimerAsRed2576592110k0.2113tetramermRFP158460730k0.25monomerJ-Red5846100.208.813dimermCherry5876100.221696monomerHcRed158861852k0.030.6dimerKatus