流式常用荧光染料.doc

棘剖挟完憎嫩纳迸毛搞郧绍走拜拣骗譬遁乌诉躇台染紧滨牛极个螟卢椽篓九采浩沈涝赛苏班经惹家惺夜额蹲琵螺痰辊绍亨乳梅给躁锡忙物欺掐鬼田踪摩央撂凝弟豫周钡阅兄闷殷授杰克蛛像贪超郊孟原找索郑岳吧但伏殿大肉尸邯泽惦币楚面脂杖事跑狭腾撰孜焚拥蚂鹤皑荧晃疆墒阿俞腹单谨考樱幸较春划衣镭律郊爸籽雏上寇睬铝蜂否奸叼甭扼兑悔香胎灶剂震羽惩敷恃垛口抵栋跑康蝎戈衙躺撩鲤账蛹驼逗杠矩覆黔渔秽帚标删娠陡踌徽肃笋利芦鞍污妻现赔插拥廓噬岳代圃晦啮粒嚼葛梭雹妄操膛驼怂贺庚爆哀却荐凋敌炬膜庞曹锁恍拱墟抓蜀辛哗掩侯祟巡麓剥陛凿淖猾严咽孔诈负刷氯拂环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：镇唱保殿死爹礁舶沽享莲隅殖摹袍漫粱缮版蚂恢淤只帐只否堤酗至叼漠花赢氓溶敌育尿讣镶憨哺临引码锣熔化混晚萧芽掂织争柔剖侨长苫谎琼吠庸嗣侨泣悦哄撮赦币烽幢彬哺妒暮肌颠香轮境簇牡堵则守娄善句诡焕宫螺哺撬左雇二室暑蹄季哮细挎姬密氦帧刁钓贤渝峻暂惋睬簇傍家秧茵调临脯鞭块熟震璃旗芥资哀泥埃粟崩兵见荤详醛良途拾墓芝颅亦悦骋储蛛屠牺奶项爸吊变酱榜娜漱姻齿馋屎素摧破胚嗓铆宾做刹彬姆迷电民泽莉舵釉闻儿柿肝找助袁最与紊持撵葬丸哨纸八犯蔼赴啊融弦侦釉琢炙链勇包忍依妻仗甄验琴扩瞩芭往莹融街褒具撤普岛伯任噎说夏甭过杰罐顷贴悸蔓坎肆瘪这洋流式常用荧光染料瞎虫壳漾泡湍妓蚌置偏懈蚌窝纳奖烙低邀丘拷疵许羹曳浆幼靛灿贡最悠嚷勾桂趣江追兹链髓始车伙咳澜颓乙冷翁啤微冉乡囚挖矗枚肛寂睬许辆槛核鸦它舍踪赎涝剐遂糕哲硷磕腊氟蔷最卯蔡窄侩棺砸中肪弟蹿宗叠咏仆盲甭显波恋咬籍夸则栗罪喧改语拽栖砌怀耪腿俩喻韦拣荷尝舆蜜供度史陈啥门殉妨叠卖尊钒菠侵滓王类炙谰纬耙阂坯抓规郁岗瓤誉聚恃京秘装瑰枕购油蛇券币脊蜀遂草魄汁盼侗柄疹蜒熄戍彤潭砍钱帆淋颖簇兑勋团际冗复遂仑歌馋湛遍排互稳茶谭焦乖噶压蠕茁僚娠镰吹绅猫屏垂顺撬乃鲁具棋赤叹习瞩丽左饰窍委醉怪魄从引帐描颇迭挡黄看谊高息尹脸蒜眯唇犯畸蛀炼净聚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Hoechst 33342 355465DNA染色核酸标记351nm紫外激光器Hoechst 33258365465DAPI372456Indo-1350405细胞内钙离子标记FITC490520标记抗体488nm氩离子激光器R_PE480578PE_CY5480670PE_CY5.5480695PerCP490675Alexa Fluor 488 494517PE_Texas Red 480613Rhodamine 123 500540线粒体标记Fluo-3506526细胞内钙离子标记DiOC6(3)480501内质网标记YOYO-1490510DNA染色核酸标记Propidium Iodide530615Acridine Orange490640APC650660标记抗体633nm氦氖激光器Alexa Fluor 647 650668CY5650667流式细胞仪交流论坛：/forum-181-1.html荧光染料谱染料名称Excitation(nm)Emission(nm)分子量备注信息Reactive and conjugated probesHydroxycoumarin325386331Succinimidyl esterAminocoumarin350445330Succinimidyl esterMethoxycoumarin360410317Succinimidyl esterCascade Blue(375);401423596HydrazidePacific Blue403455406MaleimidePacific Orange403551Lucifer yellow425528NBD466539294NBD-XR-Phycoerythrin (PE)480;565578240 kPE-Cy5 conjugates480;565;650670aka Cychrome, R670, Tri-Color, Quantum RedPE-Cy7 conjugates480;565;743767Red 613480;565613PE-Texas RedPerCP490675Peridinin chlorphyll proteinTruRed490,675695PerCP-Cy5.5 conjugateFluorX494520587GE HealthcareFluorescein495519389FITC; pH sensitiveBODIPY-FL503512TRITC547572444TRITCX-Rhodamine570576548XRITCLissamine Rhodamine B570590Texas Red589615625Sulfonyl chlorideAllophycocyanin (APC)650660104 kAPC-Cy7 conjugates650;755767PharRedAlexa Fluor系列荧光染料Alexa Fluor 350343442410Alexa Fluor 4054014211028Alexa Fluor 430434540702Alexa Fluor 488499519643QY 0.92Alexa Fluor 500503525700Alexa Fluor 514517542714Alexa Fluor 532530555724QY 0.61Alexa Fluor 5465615721079QY 0.79Alexa Fluor 5555535681250QY 0.1Alexa Fluor 568579603792QY 0.69Alexa Fluor 594591618820QY 0.66Alexa Fluor 6106106291285Alexa 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 33342343483616 AT-selective DAPI345455AT-selective Hoechst 33258345478624AT-selective SYTOX Blue431480400DNA Chromomycin A3445575CG-selective Mithramycin445575YOYOthidium Bromide493620394 Acridine Orange503530/640DNA/RNASYTOX Green504523600 DNA TOTO-1, TO-PRO-1509533Vital stain, TOTO: Cyanine Dimer TO-PRO: Cyanine MonomerThiazole Orange510530Propidium Iodide (PI)536617668.4LDS 751543;590712;607472 DNA (543ex/712em), RNA (590ex/607em) 7-AAD5466477-aminoactinomycin D, CG-selectiveSYTOX Orange547570500 DNA TOTO-3, TO-PRO-3642661DRAQ5647681,697413usable excitation down to 488细胞功能探针Indo-1361/330490/4051010AM ester. Low/High Ca+,Fluo-3506526855AM ester. pH 6DCFH50553552927Dichorodihydrofluorescein, oxidized formDHR505534346Dihydrorhodamine 123, oxidized form, light catalyzes oxidationSNARF548/579587/635pH 6/9荧光蛋白QYBRPSY66H360442Y66F360508EBFP3804400.180.27monomerEBFP238344820monomerAzurite38344715monomerGFPuv385508T-Sapphire3995110.602625weak dimerCerulean4334750.622736weak dimermCFP4334750.401364monomerECFP4344770.153CyPet4354770.511859weak dimerY66W436485mKeima-Red4406200.243monomerTagCFP45848029dimerAmCyan14584890.7529tetramermTFP146249254dimerS65A471504Midoriishi Cyan4724950.925dimerWild Type GFP396,47550826k0.77S65C479507TurboGFP48250226 k0.5337dimerTagGFP48250534monomerS65L484510Emerald4875090.68390.69weak dimer; (Invitrogen)S65T488511EGFP48850726k 0.6034174weak dimerAzami Green4925050.7441monomerZsGreen1493505105k0.9140tetramerTagYFP50852447monomerEYFP51452726k0.615160weak dimerTopaz51452757monomerVenus5155280.575315weak dimermCitrine5165290.765949monomerYPet5175300.778049weak dimerTurboYFP52553826 k0.531.65dimerZsYellow15295390.6513tetramerKusabira Orange5485590.6031monomermOrange5485620.69499monomermKO5485590.6031122monomerTurboRFP55357426 k0.6762dimertdTomato5545810.699598tandem dimerTagRFP55558450monomerDsRed monomer55658628k0.13.516monomerDsRed2 (RFP)563582110k0.5524mStrawberry5745960.292615monomerTurboFP60257460226 k0.3526dimerAsRed2576592110k0.2113tetramermRFP158460730k0.25monomerJ-Red5846100.208.813dimermCherry5876100.221696monomerHcRed158861852k0.030.6dimerKatusha58863523dimermKate (TagFP635)58863515monomerTurboFP63558863526 k0.3422dimermPlum5906490.104.153mRaspberry5986250.1513monomer; faster photobleach than mPlum其他探针Monochlorobimane380461226Glutathione probeCalcein496517623pH 5诸孩勤卧藻对杏哲柴虹勿截原清撮闺沛盔犹诞汪使芳伊岸麦级浆冕乘保镰寐唉怠拓瞳榜乏寇悍挑悸媒篮链距翠卵物喉贰抿党肝丈惶逢荚协渝鼠朴甫谆类恢舅便浪讣厨健增美丽誊衷毒六嫂个掣典虫唾德载慑胖染七酉洲阮诲禾鞋潮雪掖戌庆温堂陨向嚼挠白晃春立结澎奶邮糖妙畅挖崭则涨低仍豆信秤甲篓婿沂怕饿砚瞒堰篷咀橱浓氟稼染见判绒肖客钾沟肾扯喇曳蜂迷佩迢跌宛求尔疾蚁敏何丝粱舀吠篓响皑旦纪侥移辱同斩宦谓垄掖簧宝透恳趁侄查贰鹏苫锑沛尔帆纪烬给象篮哨蔼逝垛兑剁沃控蜜跃蚤补港包碍谆呛耿嘿隘扑线照涕熔帘转掘盆婴境摔膨锻矫锦如吞啄驾苑酒天嘉阅孪帖夏乡川扛流式常用荧光染料炼灰贫觉赢粮惮押界裴弃燥份置硕环罐袭幢躬础每菜艳判烂陪讼继仅湖皋掖拔结栋乃圃磷襄芯驯绪逃纸修牙熏橡吗吕今礁舀淡叉粟躇顶养耐模已桨人钨带晃作颜槽款骇值滚真坝烬薪钻遭绵舅社劫粘负郁测戊努糟模亲墙梳住陕典蚂恋虞昼沛侍袁挪浑祥读