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企业动态

流式细胞术常见经典问题问答!
发布时间:2018-12-20   点击次数:588次

1、LIUSHIXIBAOYISHANGDEFL2-W FL2-A FL2-H FENBIESHIZUOSHENMEDE?

彩搜网FL2-WSHIZHIJIANCEYINGGUANGDEMAICHONGKUANDU, FL2-HSHIZHIMAICHONGGAODU。TONGCHANGZAIZUOXIBAOZHOUQIFENXISHIYINGYONG,YONGYUQUCHUZHANLIANXIBAO。

彩搜网2、LIUSHITONGXINGDUIZHAOZENYANGXUANZE?

彩搜网TONGXINGDUIZHAO(Isotype Control):SHIYONGYUYIKANGXIANGTONGZHONGSHULAIYUAN、XIANGTONGYAXING、XIANGTONGJILIANGHEXIANGTONGDEMIANYIQIUDANBAIJIYAXINGDEMIANYIQIUDANBAI,YONGYUXIAOCHUYOUYUKANGTIFEITEYIXINGJIEHEDAOXIBAOBIAOMIANERCHANSHENGDEBEIJINGRANSE。RUGUOYIKANGSHIDUOKANG,KEYIYONGan normal serum(YUYIKANGXIANGTONGDEZHENGCHANGXUEQING) (must be the same species as primary antibody)。This control is easy to achieve and can be used routinely in immunohistochemical staining.ZHEIGEKEYIZIXUNSHIJISHANG。TONGXINGDUIZHAOWEIMIANYIYINGGUANGBIAOJIZHONGDEYINXINGDUIZHAO。YOUYUYINGGUANGBIAOJIDANKANGDEZULAIYUANBUTONG,YINGXUANYONGXIANGTONGLAIYUANDEWEIBIAOJIDANKANGZUOWEITONGXINGDUIZHAOLAIDIAOZHENGBEIJINGRANSE。JUGELIZI:BIRUJIANCEYIKANGWEIDANKANGDEmouse anti rat CD11b,clone OX-42 purified IgG,NEIMETADEisotype SHImouse IgG2a, SUOYIKEYIYONGpurified(CHUNHUADE) mouse IgG2aLAIZUOOX-42DETONGXINGDUIZHAO(Isotype Control)。YIBANDEDESHENGWUJISHUGONGSIHEGUONEIDEDAILIDOUYOUCHUSHOU。

TONGXINGDUIZHAO:SHIZHIYUMoAbXIANGTONGDE、WEIMIANYIXIAOSHUDEMIANYIQIUDANBAIYALEI,RUOSHIYONGZHIJIEMIANYIYINGGUANGRANSEFA,TONGXINGDUIZHAOYEYINGBIAOJIYINGGUANGSESU,RUIgG1 FITC、IgG2a、PEDENG。ZHUYAOKAOLVLEXIBAODEZIFAYINGGUANG、FCSHOUTIJIEDAODEKANGTIJIEHEHEFEITEYIXINGKANGTIJIEHEDENGYINGXIANGYINSU。CIWAI,TONGXINGDUIZHAOYUMoAbSUOBIAOJIDEYINGGUANGSESU、NONGDU、F:PBIZHI(BIAOJIDEYINGGUANGSESUYUMIANYIQIUDANBAIFENZIDEBIZHI)YINGGAIXIANGTONGWEIzuiJIA,ZHEIDUIZHUNQUESHEDINGYINXINGYUYANGXINGXIBAODEJIEBIAOYOUZHONGYAOYIYI,QIEJISHIYONGYUMoAbBUXIANGPIPEIDETONGXINGDUIZHAO,zuiHAOWEITONGYISHIYANSHI、CAIYONGXIANGTONGGONGYIHUOFANGFAZHIBEI(RUTONGYIPINPAI)DECHANPIN。

3、LIUSHIXIBAOJISHUCEDINGXIBAONEIYOULIGAINONGDUWEIHEYAOSHIYONGLVHUAGAI.ZAIWENXIANJIQITAZHUANYEWANGzhanZHONGDOUYOUCEDINGYOULIGAIDEFANGFA,JUTIRUXIA:

(1) GAIYINGGUANGTANZHENFUZAI;

(2)SUIJICEDINGMEIGUANXIBAOXUANYEYANGPIN(YIXIAJIANCHENGYANGPIN)DEDANGEXIBAODEYINGGUANGQIANGDU,JIWEIFZHI。

彩搜网(3)JIARU10%Triton X 100,(POMOYONG);JIARU1 mmol/LLVHUAGAI,(WENTISUOZAI),CHUIDAJUNYUN,FUYU1~10min,CEDINGYINGGUANGJIFmaxZHI。

彩搜网(4)JIARUEGTA,20%26mu;l(GAIAOHEJI),FUYU1~10min,JIFABOZHANG488nmCEDINGYINGGUANG,JIFminZHI。

彩搜网ZHEIGEGUOCHENGZHONGDELVHUAGAIDEZUOYONGRUHE, QINGGAORENZHIDIAN, XIEXIE。

YINWEIZHENGCHANGXUEJIANGZHONGCa2+NONGDUSHI1mM,XIBAOWAIYEDECa2+DUIXIBAONEICa2+YOUYINGXIANG。JIA0.1%tritonSHIZENGJIAMOTONGTOUXING,SHIXIBAOWAICa2+JINRUXIBAONEI,ZUOWEIDAZHI。JIAEGTASHIWEILEAOHECa2+,ZUOWEIXIAOZHI.SHENGLIHUANJINGZHONGXIBAONEIGAILIZINONGDUYUANDIYUXIBAOWAIYE(DAYUE1:10000),XIBAOMODUIGAIDETONGTOUXINGBIANHUADUIXIBAONEIGAIYINGXIANGHENDA。

彩搜网4、LIUSHIYUwersternDEQUBIE,ZHIDAOYIXIE,BUZUZHICHUQINGDAJIABUCHONG:

彩搜网(1)Western SHIJIANCEDANBAIBIAODADEJINBIAOZHUN,KEYONGYUJIANCEXIBAODUOZHONGLEIXINGDEDANBAI;LIUSHIXIBAOSHUDEFANGFADUOYONGYUJIANCEXIBAOMODANBAI,SUIRANYEKETONGGUOTriton-X100JIXIBAODAKONGDEFANGFALAIJIANCEBAONEIDANBAI,DANDUIYUFENMIDANBAIQUESHIWUNENGWEILIDE;

彩搜网(2)WesternCEDANBAIHANLIANGDUIKANGTIDELIANGXUYAOHENSHAO,YIBAN1:1000ZUOYOU,ERLIUSHIDUIKANGTIDEXUYAOLIANGHENDA,YIBAN1:50(HENLANGFEIKANGTI);

彩搜网(3)CAIYONGWesternFANGFAJIANCEXIBAODANBAIHANLIANGDEBIANHUAYIBANYAOYOUNEICAN,ERLIUSHIYIBANYAOBAMEIGEYANGPINFENWEILIANGFEN,TONGSHIDOUZUOZHIJIAERKANG(FITCBIAOJIDE)DEDUIZHAO,ZHEIYANGPINGJUNDAOMEIGEXIBAODEFAGUANGJIUBUYONGNEICANLE;

(4)JIUGERENJINGYANERYAN,LIUSHIYONGDUOKANGBUHAO,DANKANGBIAOCHULAIBUCUO。

彩搜网BUGUOLIUSHIDEYINGYONGYUANLIZHUYAOHAISHIKANGYUANHEKANGTIFANYINGHEwestern、MIANYIZUHUAMEIYOUBENZHICHABIE,DANSHIYOUYUMIANYIZUHUAHEwesternDOUYOUMEICUIHUADIWUDEFANGDATIXI,YINCI,LIUSHIBUSHIHEJIANCEDIBIAODADEDANBAI,DIBIAODADEHAISHICAIYONGwestern(YOUQISHIHUAXUEFAGUANGDIWUGENGJIA)HEMIANYIZUHUAGENGHAO。DANGRAN,LIUSHIDADEYIGETEDIANJIUSHI,KEYIDINGLIANG(BAIFENBI),RUGUOSHIGAOBIAODADEYONGLIUSHIXIBAOYIFEICHANGYOUYOUSHI。

5、FL1, FL2, FL3 DEQUBIESHISHENME? SHIZHIZAIBUTONGWEIZHICEDEDEYINGGUANG?HAISHIBUTONGBOZHANGDEYINGGUANG?ZHEI3GECANSHUDAODICEDESHISHENME?YOUSHENMEYIYI?

NIDELIJIESHIZHENGQUEDE,QISHIFL1, FL2, FL3 SHIZHIBUTONGDEYINGGUANGTONGDAO.JUGELIZILAISHUOBA.WOMENYONGFITC/PESHUANGBIAOJIDEXIBAO,ZAI488nmDEJIGUANGJIFAXIA,ZHEILIANGZHONGYINGGUANGRANLIAOFENBIEFACHUBOZHANGBUTONGDELVSEHECHENGSEYINGGUANG,RANHOUZHEIFACHUDEYINGGUANGZAITONGGUOLVGUANGPIANFENKAI,DUIYINGDESHIBUTONGDEBOZHANGDELVGUANGPIAN,YIBANZAIfl1NEIDESHI530nmDELVGUANGPIAN,ZAIFL2NEIDESHI575nmDELVGUANGPIAN,ZHEIYANGJIUKEYIBAZAIYIQIDEYINGGUANGFENKAILE。

6、YONGYUZUOWestern HUOELISADEYIKANGKEFOUYONGYULIUSHIXIBAOSHU?

KANNISHUOMINGSHUSHANGSHIZENMESHUODELE,RUGUOMEIYOUSHUOJIUBUKEYIZUOLIUSHI,XUYAOLINGWAIMAILE。

彩搜网7、MFIHEGFIDEYIYI?

Geo Mean,JIAOZUOJIHEJUNSHU(geometric mean),CHANGYONGYUDENGJIZILIAOHEDUISHUZHENGTAIFENBUZILIAO,HECHANGYONGDESUANSHUJUNSHU(mean)DEJISUANFANGFABUTONG。%26ldquo; %totalHUOZHE%gateDEJIEGUO%26rdquo;DAIBIAODESHIBAIFENLV,JIXIBAOZHANZONGTIXIBAOHUOZHESHIMENNEIXIBAODEBAIFENBI;YONGBAIFENBIZUOWEITONGJIZHIBIAO,SHIYONGYUYINGGUANGBIAODAJIAOQIANGDEZHIBIAO。WOKANDAONIDELIUSHITUSHANG,JIANCEYANGBENDEYINGGUANGQIANGDUBUSHIHENQIANG,SHUYURUOBIAODADEZHIBIAO,RUOYONGBAIFENLVTONGJI,JIUSHIHUISHIQUYIBUFENRUOYANGXINGDESHUJU。WOJIANYIKEYIYONGPINGJUNYINGGUANGQIANGDU(YEJIUSHImeanZHI)ZUOWEITONGJIZHIBIAO,BIJIAOYINGGUANGQIANGDUDEBIANHUA。

彩搜网8、LIUSHIJISHUZHONGDEAPC,pure BIAOJISHISHENMEYISINI?

YINGGUANGWUZHITONGCHANGSHIZHINEIXIEZAISHOUDAOYIGEBOZHANGJIFAHOU,CHUYUYIGENENGLIANGBUWENDINGDEZHUANGTAI,NENGFASHEYIGEBOZHANGBIJIFABOZHANGZHANGDEGUANGDEYILEIWUZHI。DANGRANYINGGUANGBINGBUDOUSHISHOUJIFAHOUFASHEDE,RUYIXIESHENGWUKEYIFASHEYINGGUANG,RUYINGHUOCHONG,FAGUANGXIJUNDENG,TAMENFACHUDEGUANGSHITONGGUOTINEIDEMEICUFANYINGERCHANSHENGDE,ZHEIGEMEITONGCHANGBEICHENGWEIYINGGUANGSUMEI.EBSHIYIZHONGYINGGUANGWUZHI,TAZAISHOUDAOZIWAIXIANZHAOSHEHOUKEYIFACHUYIKEJIANGUANG,CHANGYONGYUDNA、RNADIANYONGZHONG。

APC

YINGWENQUANMING:allophycocyanin;ZHONGWENMING:BIEZAOQINGDANBAI;DAXISHOUFENG:650nm,DAFASHEYINGGUANGFENG:660nm,SHIYONGYUSHUANGJIGUANGLIUSHIYI,KEBEI600-640nmBOZHANGDEJIGUANGJIFA。

PerCP

彩搜网YINGWENQUANMING:peridinin chlorophyll protein,ZHONGWENMING:DUOJIAZAOYELVSUDANBAI;DAJIFABOFENG490nm,BEI488nmDEYALIZIJIGUANGJIFAHOU,FASHEGUANGFENGZHIYUEWEI677nm,YUFITC、PEJINXINGDUOSEYINGGUANGRANSEBUCHANGZHONGDIEJIAOSHAO,FEITEYIXINGJIEHEYEJIAOSHAO。SUIRANJIAOYISHIYONG,DANLIANGZICHANLIANGBUGAO,DUOYONGYUJIANCEBIAODAJIAOGAODEKANGYUAN。

9、ZENYANGYONGLIUSHIXIBAOYILAIJIANDINGXIAOSHUBMDC?

彩搜网JIANCEXIAOSHUBMDCZHUYAOSHICONGXINGTAIXUEHEXIBAOBIAOMIANFENZILIANGFANGMIANLAIJIANDINGWOZUODESHIHOUJIUZUOLEPUTONGGUANGJINGXIAXINGTAI、DIANJING(SAOMIAOHETOUSHE),LINGWAIJIANCELEXIAOSHUBMDCBIAOMIANDEGONGCIJIFENZIRUCD86 、CD11、CD80。HAIZUOLEMHC IILEIFENZIDEJIANCE,HAIYOUMHCILEIFENZIDEJIANCE,ZHEIXIEFENZIZAIDCBIAOMIANDOUBUSHITEYICUNZAIDE,ZAIJUSHIXIBAO、LINBAXIBAOBIAOMIANYEYOUBIAODA,SUOYIMUQIANBINGMEIYOUFEICHANGTEYIXINGDEFANGFAJIANCENIDEXIBAOYIDINGJIUSHIDC,DANSHICONGXINGTAIHEBIAOMIANFENZIDEJIANCEJIEHELAIKAN,XIAOGUOJIUBIJIAOHAOLE.YIBANZAIYOUZHIDEDCSHANGSHUFENZIBIAODASHUIPINGJIAODI,DCCHENGSHUGUOCHENGZHONG,SHANGSHUFENZIBIAODACHENGSHANGDIAOQUSHI,NIKEYIZAIBUTONGDEPEIYANGSHIJIANFENBIEJIANCE。

10、QINGWENLIUSHIXIBAOSHUDANKANGZHIJIEYINGGUANGXUYAOJIZHONGDUIZHAO?

SHOUXIANQUERENNIYONGDEZHIBIAOKANGTIDEYINGGUANGRANLIAOSHISHENME,ZAIQUEDINGGAIKANGTIDELAIYUANZHONGSHU,XUANZEXIANGYINGDETONGXINGDUIZHAO。RU:NIXIANZAIXIANGJIANCEXIAOSHULINBAXIBAOBIAOMIANDECD4KANGYUAN, YINGGUANGRANLIAOWEIFITC, KANGTILAIYUANWEIDASHUDEIgG1YAXING, JIRat anti Mouse CD4-FITC (IgG1), NISUOXUYAODETONGXINGDUIZHAOJIUYINGGAISHIRat IgG1-FITC, YIBANDEKANGTISHUOMINGSHANGDOUHUIXIEQING。

11、6KONGBANDEXIBAOLIANGNENGFOUZUOLIUSHIYA?

彩搜网DUIYU6KONGBANERYAN,MEIKONGDEBIAOMIANJIWEI10 cm2,YIXIBAOZHONGLEIBUTONGZAIZHANGMANSHIDADAODESHULIANGCONG5*10DE5CIFANGDAO5*10DE6CIFANGBUDENG。6KONGBANMEIWENTIDE!SHENZHI24KONGBANYEKEYIZHENGCHANGZUO。BUGUOYONGYUDIAOSHIYIQICANSHUDEZHENGCHANGXIBAOYAODUOZHUNBEIYIXIE。

彩搜网12、XUEYELIDEmononuclear cell(CHUWAIHONGXIBAO,XUEXIAOBANHEPOSUIDEXIBAOSUIPIAN),NENGBUNENGYONGXIBAODAXIAOLAIgating,ZHIKANWOGUANXINDEXIBAO?

(1)HONGXIBAOHAISHIXIAOYIXIE,WUFA100%QUFEN,DANHAISHIKEYONGBADABUFENHONGXIBAOgateDIAO。

彩搜网(2)RONGXUEYEHENZHONGYAO,RUGUONIDEHONGXIBAODUODEHUA。(WOCAIBUSHAO,RUGUOYONGficolDEHUA)。KEYIYONGLVHUAANRONGYE,RUACK(Lonzo),

(3)CD45SHISUOYOUBAIXIBAODOUBIAODADE,KEYIYONGLAIQUFENRBC vs. WBC

13、XIBAOMODANBAIBIANHUASHIYONGLIUSHIJIANCEHAO,HAISHIYAOZUOwestern?

MODANBAIDETIQU,HAOXIANGHENFEIGONGFU,TIQUDEBUHAO,westernJIEGUOYEJIUBUKEXIN。LIUSHIXUYAODEKANGTIHENSHAO,LIUSHINENGGOUJIANCEYANGXINGBIAODAXIBAODEBILI,westernNENGDUIZONGDEBIAODADINGLIANG,GEYOUYOUQUEDIAN。

彩搜网14、FCMJIANCEBIAODAJIEGUODAODISHIYONGBAIFENBIHAISHIMFI。WOZUOCHUSHIDANFENG,YUANBENWOJUEDEYONGMFIBIAOSHIJIAOHAO。DANWOZUODE2GEDANBAIHENQIGUAI,YIGEBIAODALVGAO,DANMFIZHIDI;LINGYIGEBIAODALVDI,DANMFIZHIGAO。WOYOUZUOLESYBR GREEN DINGLIANGPCR,FAXIANmRNA DEBIAODAJIBENYUBAIFENBIXIANGFU,ERYUMFIZHIBUTAIYIZHI。

GERENRENWEIRUGUOYOUMINGXIANYINXINGXIBAOQUNHEYANGXINGXIBAOQUN,YINGJISUANBAIFENBI;WUMINGXIANFENQUN,JINYINGGUANGQIANGDUFASHENGBIANHUADEYINGGAIYONGMFI。RUGUOSHIZHENGTIFENGYI(HUOZHEJIAODANFENG),NEIMEGENBENBUCUNZAIMFIZHIWAIDERENHEHELIDEZHIBIAO,BUGUANMFIGENQITAZHIBIAOWENHEDURUHE,ZHEIJIUSHIKEGUANSHUJU、KEGUANSHISHI。

15、PEIYANGXIBAODEbcl-2JIBaxDANBAIDEJIANCE?

SHOUXIANZHICHENGDANXIBAOXUANYE,PBSXIDIHOUYONGBAONEIRANSESHIJIHE(HENDUOGONGSIDOUYOU,QISHIJIUSHIGUDINGJIJIAZENGTOU/DAKONGJI)JINXINGCHULI,ZAIJINXINGKANGTIFUYUBIAOJIRANSE,XIDIHOUSHANGYANG。

16、LIUSHIJIANCEBAONEIKANGYUANSHIDAKONGYEDEPEIFANG?

DAKONGYEYOUHENDUOZHONG,FANGFAYEYOUHENDUO。YUNIDESHIYANFANGFAXIANGGUAN。WOYONGGUOJIUJINGGUDINGHUOTriton X-100(WOZUODEDOUSHIPEIYANGDEXIBAO):

(1)70%DEJIUJINGGUDING24XIAOSHIJIKE,BUZAIXUYAOZAIDAKONG。RUZAIPIRANSECEXIBAOZHOUQIHUODIAOWANG。

(2)Triton X-100SHIBIJIAOQIANGLIEDECHUANKONGJI。0.1% Triton X-100/PBS(ZAIJIA0.1%BSA) SHIBIJIAOWENHEYIDIAN。NONGDUKESHIDANGTIGAO。ZUOYONGSHIJIANYIJIFENZHONGWEIYI。ZHEIGESHIJIANBIXUYAOZIJISHI。SHIJIANZHANGLEXIBAOYIPOLIE,DUANLEZEDAKONGBUGOU。RUGUONIYAORANBAOJIANG,ZESHIJIANSHIDANGDUANXIE,RUGUOYAORANNEIZHIWANGHUOHENEIDENG,YOUYUYAODUILIANGCENGMOXINGJIEGOUDAKONG,SHIJIANDANGRANHUIZHANGYIXIE

(3)KEYISHISHI0.1% saponin(ZAOSU)

彩搜网17、BIAOBENBIXUZAIKANGTIRANSEHOU30FENNEIJIANCEMA?

SOPSHIZHEIMESHUODE:

彩搜网(1)Keep samples after staining covered with aluminum foil and at 4 0C (in the refrigerator) until they are run for flow analysis. The samples should be analyzed within 48 hours. Fluorescence loses its brightness if cells are not kept properly: not at 4 0C, or are exposed to light

(2)If cells are not fixed with paraformaldehyde, keep the samples on ice and run them immediately after staining is accomplished

彩搜网YINCI,RUGUONIMEIYOUYONGPBSXIDEHUA,KEYIYONGparaformaldehydeGUDING,FANGZHI48XIAOSHI。XILEHOUYINGGAIJINKUAIJIANCE。RUGUOZUOHAOBUNENGJISHIJIANCE,WOMENYIBANSHIJIARUDENGLIANGDE2%DEDUOJUJIAQUANGUDING,4DUBINGXIANGBIGUANGZHUCUN。YIBANGUOYEMEIYOUWENTI,DIERTIANTONGYANGPBS 2ml XIDIXIBAOYICI,SHANGJIJIANCE。

彩搜网18、RUHEFENXUANYUANSHIHONGXIBAO?

CD71(ZHUANTIEDANBAISHOUTI)--YOUZHIXIBAODEBIAOJI

CD235(GPA)--HONGXIBAODEBIAOJI(XIAOSHUYOUTer119)

彩搜网SHUANGYANGXINGXIBAOJIWEIYOUZHIHONGXIBAO,DANWUFAQUFENYUANSHIJIZHONGWANYOUHONG.

19、QINGWENLIUSHIJIANCEMODANBAIBIAODADEBIANHUAYONGDUOKELONGKANGTICHUJIEGUODEJIHUIDAMA?

LIUSHIDEKANGTIHEWBDEKANGTISHIBUTONGDE(YOUBUFENKEYITONGYONG),DUOKANGYIBANSHIDADEBIANXINGDANBAIWEIKANGYUANZHIBEIDE,ERWBDEZHONGBEIJIANCEDANBAIJIUSHIBIANXINGZHUANGTAI,SUOYIHENWENHE,ERLIUSHIZHONGYIBANDANBAIHAISHIBAOCHILETIANRANGOUXIANG,SUOYICHANSHENGDEDUOKANGKENENGBUTAIHAO,ERXUYAOYONGHENXIAOTEDINGDEJUEDINGCUWEIKANGYUANZHIBEIDANKELONGKANGTI。

彩搜网20、ZAIXIAJINZUORENWAIZHOUXUELIUSHI,ZUOBIAOMIANJIQIBAONEIRANSE,SHIJISHUOMINGSHUSHUOYAOXIANFENLIWAIZHOUXUEDANGEHEXIBAOZAIRANSE,WOYOUSHIHOUFENDEHONGXIBAOBIJIAODUO,QITADEFANGFASHIGUOLE,DOUGAIJINDEBUHAO,WODANXINHONGXIBAOHUIYOUYINGXIANG,XIANGZAIFENLICHUPBMCYIHOURUGUOHONGXIBAOBIJIAODUODEHUA,JIUYONGHONGXIBAOLIEJIEYELIEJIEYIXIA,DANSHIWOYOURUXIAWENTI,XIWANGZHEIYANGZUOGUODETONGzhiZHIDIANYIXIA,GANJIBUJIN。

(1)YOURENZHEIYANGZAIFENLIPBMCYIHOUZAILIEJIEDEMA(WOZHIDAOYIBANSHIZAIQUANXUERANSEHOUZAILIEJIEHONGXIBAO)?

(2)ZHEIYANGHUIYINGXIANGLIUSHIJIANCEMA?

(3)YONGDELIEJIEYEPEIFANGSHISHENME(zuiHAOSHIZIJIYONGGUODE)。

(4)YONGFASHISHENME{WOSHIZAIRUGUOPBMCFENCHULAIYIHOUYAOSHIHONGXIBAOBIJIAODUODESHIHOUSHIYONG,ZHEIYANGYAOZENYANGBAFENCHUDEPBMCXISHI,JIADUOSHAOLIEJIEYE LIEJIEDUOZHANGSHIJIAN,LIEJIEHOUXIDIJICIDENGDENG}。

WOCENGZUOGUSUIYEFENLIDANGEHEXIBAOERHOUZUOLIUSHI,YIBANLAISHUOYONGFicollFENGUOZHIHOUJIBIANHAIYOUHONGXIBAOCANLIU,SHANGLIUSHIYEKETONGGUO%26ldquo;SHEMEN%26rdquo;GENJUXIBAOXINGTAIDAXIAOJIANGHONGXIBAOTICHU。RUOHONGXIBAOCANLIUTAIDUO,ZEKETONGGUOHONGXIBAOLIEJIEYEJINYIBUQUCHUZHI。WOYONGDELIEJIEYEPEIFANGSHI:

TANSUANQINGJIA(KHCO3) 1.0g

LVHUAAN(NH4CL) 8.3g

EDTA-Na2 0.037g JIASHUANGH2OZHI1000ml,WEIGONGZUOYE。

彩搜网PEIHAOHOU4DUBINGXIANGBAOCUN,SHIYONGSHIXIAOGUOGENGHAO。WOSHIZAIFicollFENGUOZHIHOUYONGDE,SUOYIYIBANGENJULIXINHOUEPGUANLIXIBAOTUANDEDAXIAOJIAHONGXIBAOLIEJIEYE,TONGCHANG5mlGUSUIYEFENLIDANGEHEXIBAOHOUDEDAODEXIBAOZAIJIA500ulLIEJIEYE,ZHENDANGHUNYUNZHI2-5FENZHONGHONGXIBAOJIUJIBENSHANGLIEJIEGANJINGLE。XIDICISHUKANBAIXIBAODUOSHAOERDING,YINWEIXIDECISHUYUEDUO,SUNSHIJIUYUEDUO。WOYIBANXIYIDAOLIANGCIJIUOKLE.ZUOLEHENJIUMEIFAXIANZHEIYANGCHULIYINGXIANGJIEGUO。YOUYIDIAN,WOSHIZAIZHEIYANGCHULIHOUCAIBIAOJIKANGTIDE,LOUSHANGDESHUOSHIBIAOJIGUOCAIRONGHONGXIBAO,SUOYIJIANYIZAIRONGDESHIHOUSHIJIANBUYIGUOZHANG,XIDICISHUYEBUYIGUODUO,YIMIANJIANGBIAOJISHANGDEDANKANGXITUO。

21、QINGWENXIABIAOZHONGLIUSHIXIBAOYIJICHUDEPINGJUNZHISHISHENMEYISI,SHIZENYANGDEDAODE,YOUXIEWENXIANZHONGTIDAO%26ldquo;fluorescence per cell%26quot;,SHIXIAMIANDEmeanZHIMA,HAISHIYAOYONGmeanZHICHUYIDI1LANDEXIBAOSHUevents,CAINENGDEDAOfluorescence per cell ?

mean ZHIJIUSHIMEIGEXIBAODEPINGJUNYINGGUANGQIANGDU,BUYONGZAICHUXIBAOSHU。ZHEIZHISHIGEXIANGDUIZHI,RUGUOQIUZHI,YINGYONGYIZHIYINGGUANGJINONGDUZHIDUIYINGYIQIJICHUDEmeanZHI,ZUOBIAOZHUNQUXIAN。YIHOUNIDEDAODEmeanZHI,JIUKEYIGENJUZHEITIAOBIAOZHUNQUXIANCHACHUZHENZHENGDEYINGGUANGNONGDUZHI。

22、LIUSHIZHONGJIANCEXIBAOBIAOMIANHEXIBAONEIDANBAIBIAODASHUIPINGDEKANGTIYOUHEQUBIE?

彩搜网KANGTIZHISHIZHENDUIXIANGYINGKANGYUANDE,ZHIYUSHIZHENDUIBAONEIHAISHIBAOMODUINIJIANCELAISHUOBINGWUMINGXIANYINGXIANG,NIZHIXUYAOCHAQINGCHUDAODISHIZHENDUINAYIGEBUWEIDEKANGYUANDE,YINGGAISHIKEYIYONGTONGYIZHONGKANGTIJINXINGJIANCEDE,ZAILIUSHICAOZUOSHIZHIXUZHUYI:RUGUOSHIZHENDUIBAONEI,ZEXUYAOJIAPOMOJI,RANGKANGTINENGHEXIANGYINGKANGYUANJIECHU,FOUZEJIUBUXUYAOLE。

23、QINGJIAOGEWEIQIANBEI:ZHIYAOSHIYINGGUANGKANGTIJIUKEYIYONGYUGONGJUJIAOXIANWEIJINGMA?

彩搜网YIBANQINGKUANGXIADOUSHIKEYIDE。DANYOUSHIHOUQIANGDUBUGOU,XUYAOXUANZEYINGGUANGQIANGDUDADERANLIAO,BIRUAlexaXILIEDE。

彩搜网24、LIUSHIXIBAOYIJIANCEXIBAOZHOUQISHIFOUXUYAOJIHONGXIBAOZUOCANZHAO?

BUYONG,BIAOZHUNWEIQIUZUOWANlaser alignment(GUANGLUXIAOZHENG)JIUKEYILE。JINLIANGBAWEIQIUDEGESECVZHIKONGZHIZAI1.5YINEI。

彩搜网25、WOXIANZAIYAOYONGJIANJIEFALIUSHIXIBAOSHU,GANGMAILEYINGGUANGERKANG,SHIKPLDEfluorescein-labeled affinity purified antibody to mouse IgG+IgM(H+L) produced in goat .SHUOMINGSHUOSHANGTISHIYAOYONGTBSHUOSHIBSAHUOmilk diluent/blocking solutionYEXISHI。XISHIZHI1:10~1:100。QINGWENBSASHIBUSHIXIAONIUXUEQING,YAOYONGDUOSHAONONGDUDE。TBSYEWOMEIYOU,HAIYOUKEFOUYONGZHUSHEYONGSHUIXISHI。

BSA:NIUXUEQINGBAIDANBAI,NONGDUKEYIZAIYIDINGFANWEINEI1%~5%,JUTIKECANKAOQITAKANGTISHUOMINGSHU。

TBS:Tris buffer saline。JIUSHITrisDEYANRONGYE,HENCHANGGUIDERONGYE,PEIFANG:20mM Tris-HCl(ph8.0),150mM NaCl。

彩搜网NISHUODEZHUSHEYONGSHUIYINGGAIJIUSHISHENGLIYANSHUIBA,ZHEIDUIKANGTILAISHUOSHIBULIDE,SUIRANZAIJICHUTIAOJIANHENCHADEQINGKUANGXIAHUIYONGZHEIGERONGYE,DANSHIHAIBURUPBSRONGYE。JIRANSHIZUOLIUSHI,ANZHAOZHENGGUICHENGXUCAOZUOKANGTI,DEDAODEJIEGUOYENENGFANYINGZHENshiQINGKUANG。

26、LIUSHIXIBAOJIANCEZHONGZENYANGBADUIZHAOHEYANGBENDEJIANCEJIEGUOFANGZAIYIZHANGTUZHONG?

FENXIHUOQUSHUJUSHIFENKAIJINXINGDE,BUKEYIHUNZAIYIQISHANGYANG。SHOUXIANTONGGUOYINXINGDUIZHAODIAOJIEJIBENDIANYA,GUDINGTIAOJIANHOUJINXINGYANGPINJIANCE,FENBIEBAOCUNJIEGUO。LIYONGLIUSHIRUANJIANDEmultigraphZHONGDEoverlapKEYIJIANGYINXINGDUIZHAOHEYANGPINJIEGUOXIANGYINGTUJINXINGDIEJIA,ZHEIZHISHIHUOQUSHUJUHOUDUIJIEGUODEZUHEHEJIAGONG。

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