Lub Countstar system muab cov duab cytometer thiab cov txee ntawm tes rau hauv ib lub rooj ntev zaum saum toj kawg nkaus.Daim ntawv thov no-tsav, compact, thiab automated cell imaging system muab ib qho kev daws teeb meem tag nrho rau kev tshawb fawb qog noj ntshav, suav nrog kev suav ntawm tes, kev muaj peev xwm (AO / PI, trypan xiav), apoptosis (Annexin V-FITC / PI), cell Lub voj voog (PI), thiab GFP / RFP hloov pauv.
Abstract
Mob qog noj ntshav yog ib qho tseem ceeb ntawm kev tuag thoob ntiaj teb, thiab kev txhim kho cov kev kho mob qog noj ntshav tshiab yog qhov tseem ceeb heev.Cov kab mob qog noj ntshav yog qhov kev tshawb fawb tseem ceeb ntawm cov qog nqaij hlav, ntau cov ntaub ntawv yuav tsum tau soj ntsuam los ntawm cov qog nqaij hlav cancer.Qhov chaw tshawb fawb no xav tau ceev, txhim khu kev qha, yooj yim thiab cov ncauj lus kom ntxaws ntawm tes.Countstar system muab cov kev daws teeb meem yooj yim rau kev tshuaj xyuas mob qog noj ntshav.
Kawm Cancer Cell Apoptosis los ntawm Countstar Rigel
Apoptosis assay yog niaj hnub siv nyob rau hauv ntau lub chaw soj nstuam rau ntau lub hom phiaj los ntawm kev ntsuam xyuas kev noj qab haus huv ntawm cov kab lis kev cai ntawm tes mus rau kev ntsuam xyuas cov tshuaj toxicity ntawm lub vaj huam sib luag ntawm cov tebchaw.
Apoptosis assay yog ib hom siv los txiav txim qhov feem pua ntawm apoptosis ntawm cov hlwb los ntawm Annexin V-FITC / PI staining method.Annexin V khi rau phosphatidylserine (PS) nrog rau thaum ntxov apoptosis cell los yog necrosis cell.PI tsuas yog nkag mus rau necrotic / heev lig-theem apoptotic hlwb.(Daim duab 1)
A: Early apoptosis Annexin V (+), PI (-)
B: Late apoptosis Annexin V (+), PI (+)
Daim duab 1: Loj cov ntsiab lus ntawm Countstar Rigel duab (5 x magnification) ntawm 293 hlwb, kho nrog Annexin V FITC thiab PI
Cell Cycle Analysis of Cancer Cell
Lub voj voog ntawm tes lossis lub voj voog ntawm tes faib yog cov xwm txheej uas tshwm sim hauv lub xovtooj ntawm tes ua rau nws qhov kev faib tawm thiab sib npaug ntawm nws cov DNA (DNA replication) los tsim ob lub hlwb.Hauv cov hlwb nrog lub nucleus, zoo li hauv eukaryotes, lub voj voog ntawm tes kuj tau muab faib ua peb lub sijhawm: interphase, mitotic (M) theem, thiab cytokinesis.Propidium iodide (PI) yog cov tshuaj tua kab mob nuclear uas nquag siv los ntsuas lub voj voog ntawm tes.Vim tias cov dyes tsis tuaj yeem nkag mus rau cov hlwb nyob, cov hlwb raug kho nrog ethanol ua ntej staining.Tom qab ntawd tag nrho cov cell yog stained.Cells npaj rau kev faib yuav muaj ntau ntxiv ntawm DNA thiab tso saib proportionally fluorescence.Qhov sib txawv ntawm fluorescence siv los txiav txim qhov feem pua ntawm cov hlwb hauv txhua theem ntawm lub voj voog ntawm tes.Countstar tuaj yeem ntes cov duab thiab cov txiaj ntsig yuav tshwm sim hauv FCS nthuav qhia software.(Daim duab 2)
Daim duab 2: MCF-7 (A) thiab 293T (B) tau stained nrog cell cycle Detection Kit nrog PI, cov txiaj ntsig tau txiav txim los ntawm Countstar Rigel, thiab tshuaj xyuas los ntawm FCS nthuav qhia.
Viability thiab GFP Transfection Determination hauv Cell
Thaum lub sij hawm bioprocess, GFP feem ntau yog siv los fuse nrog recombinant protein raws li qhov taw qhia.Txiav txim siab GFP fluorescent tuaj yeem cuam tshuam txog lub hom phiaj ntawm cov protein.Countstar Rigel muab kev ntsuam xyuas sai thiab yooj yim rau kev kuaj GFP kev hloov pauv nrog rau kev muaj peev xwm.Cells tau stained nrog Propidium iodide (PI) thiab Hoechst 33342 los txhais cov neeg tuag ntawm tes thiab tag nrho cov cell cell.Countstar Rigel muaj txoj hauv kev ceev, ntau npaum li cas rau kev ntsuas GFP kev nthuav qhia kev ua tau zoo thiab muaj txiaj ntsig zoo tib lub sijhawm.(Daim duab 4)
Daim duab 4: Cells nyob siv Hoechst 33342 (xiav) thiab feem pua ntawm GFP qhia cov hlwb (ntsuab) tuaj yeem txiav txim tau yooj yim.Cov cell tsis tuaj yeem yog xim av nrog propidium iodide (PI; liab).
Viability thiab Cell suav
AO/PI Dual-fluoresces suav yog hom tshuaj ntsuam xyuas siv los kuaj xyuas cov cell concentration, kev vam meej.Nws muab faib ua cov kab ntawm tes suav thiab suav cov xov tooj tseem ceeb raws li cov xov tooj sib txawv.Cov tshuaj muaj kev sib xyaw ua ke ntawm ntsuab-fluorescent nucleic acid stain, acridine txiv kab ntxwv, thiab redfluorescent nucleic acid stain, propidium iodide.Propidium iodide yog ib daim nyias nyias dye uas tsuas yog nkag mus rau hauv cov hlwb nrog kev cuam tshuam cov membranes thaum acridine txiv kab ntxwv nkag mus rau tag nrho cov hlwb hauv cov pejxeem.Thaum ob lub dyes muaj nyob rau hauv lub nucleus, propidium iodide ua rau txo qis hauv acridine txiv kab ntxwv fluorescence los ntawm fluorescence resonance zog hloov (FRET).Raws li qhov tshwm sim, cov cell nucleated nrog cov kab mob tsis zoo stain fluorescent ntsuab thiab suav tias yog nyob, hos cov cell nucleated nrog cov kab mob cuam tshuam tsuas yog stain fluorescent liab thiab suav tias tuag thaum siv Countstar Rigel system.Cov khoom tsis muaj nucleated xws li cov qe ntshav liab, platelets thiab cov khib nyiab tsis muaj fluoresce thiab tsis quav ntsej los ntawm Countstar Rigel software.(Daim duab 5)
Daim duab 5: Countstar tau ua kom zoo dual-fluorescence staining txoj kev rau kev yooj yim, raug txiav txim siab ntawm PBMC concentration thiab muaj peev xwm.Cov qauv stained nrog AO / PI tuaj yeem txheeb xyuas nrog Counstar Rigel
