Dongosolo la Countstar limaphatikiza chithunzi cha cytometer ndi chowerengera cha cell kukhala chida chimodzi chapamwamba cha benchi.Kachitidwe kameneka kamene kamayendetsedwa, kaphatikizidwe, komanso ka makina opangira ma cell kumapereka yankho lathunthu pakufufuza kwa maselo a khansa, kuphatikiza kuwerengera ma cell, kuthekera (AO/PI, trypan blue), apoptosis (Annexin V-FITC/PI), cell cycle (PI), ndi GFP/RFP transfection.
Ndemanga
Khansara ndi imodzi mwazomwe zimayambitsa imfa padziko lonse lapansi, ndipo kupangidwa kwa njira zatsopano zothandizira khansa ndizofunikira kwambiri.Selo la khansa ndiye chinthu chofunikira pakufufuza za khansa, zidziwitso zosiyanasiyana ziyenera kuwunikiridwa kuchokera ku cell ya khansa.Dera lofufuzirali likufunika kusanthula mwachangu, kodalirika, kosavuta komanso mwatsatanetsatane.Countstar System imapereka njira yosavuta yothetsera kusanthula maselo a khansa.
Phunzirani Cancer Cell Apoptosis ndi Countstar Rigel
Mayeso a Apoptosis amagwiritsidwa ntchito pafupipafupi m'ma laboratories ambiri pazifukwa zosiyanasiyana kuyambira pakuwunika thanzi la zikhalidwe zama cell mpaka kuwunika kawopsedwe ka gulu lamagulu.
Apoptosis assay ndi mtundu womwe umagwiritsidwa ntchito pozindikira kuchuluka kwa apoptosis yama cell ndi njira yodetsa ya Annexin V-FITC/PI.Annexin V imamanga ku phosphatidylserine (PS) yokhala ndi cell yoyambirira ya apoptosis kapena necrosis cell.PI imangolowa m'maselo a necrotic/ochedwa kwambiri.(Chithunzi 1)
A: Early apoptosis Annexin V (+), PI (-)
B: Late apoptosis Annexin V (+), PI (+)
Chithunzi 1: Tsatanetsatane wa zithunzi za Countstar Rigel (5 x magnification) za maselo 293, othandizidwa ndi Annexin V FITC ndi PI.
Ma cell Cycle Analysis of Cancer Cell
Kuzungulira kwa ma cell kapena kugawikana kwa ma cell ndi mndandanda wazinthu zomwe zimachitika mu cell zomwe zimatsogolera kugawika kwake ndikubwereza kwa DNA yake (DNA replication) kupanga ma cell awiri aakazi.M'maselo okhala ndi phata, monga mu eukaryotes, kuzungulira kwa selo kumagawidwa m'magulu atatu: interphase, mitotic (M) phase, ndi cytokinesis.Propidium iodide (PI) ndi utoto wodetsa wa nyukiliya womwe umagwiritsidwa ntchito nthawi zambiri kuyeza kuzungulira kwa ma cell.Chifukwa utoto sungathe kulowa m'maselo amoyo, ma cell amakhazikika ndi ethanol asanadere.Ma cell onse amadetsedwa.Maselo omwe akukonzekera magawano adzakhala ndi kuchuluka kwa DNA ndikuwonetsa molingana ndi kuchuluka kwa fluorescence.Kusiyana kwa mphamvu ya fluorescence kumagwiritsidwa ntchito kudziwa kuchuluka kwa ma cell mu gawo lililonse la ma cell.Countstar ikhoza kujambula chithunzichi ndipo zotsatira zidzawonetsedwa mu pulogalamu ya FCS Express.(Chithunzi 2)
Chithunzi 2: MCF-7 (A) ndi 293T (B) anali odetsedwa ndi selo cycle Detection Kit ndi PI, zotsatira zake zinatsimikiziridwa ndi Countstar Rigel, ndi kufufuzidwa ndi FCS Express.
Viability ndi GFP Transfection Determination mu Cell
Panthawi ya bioprocess, GFP nthawi zambiri imagwiritsidwa ntchito kuphatikiza mapuloteni ophatikizanso ngati chizindikiro.Dziwani kuti fluorescent ya GFP imatha kuwonetsa zomwe mukufuna.Countstar Rigel imapereka kuyesa kwachangu komanso kosavuta kuyesa kufalikira kwa GFP komanso kutheka.Maselo adadetsedwa ndi Propidium iodide (PI) ndi Hoechst 33342 kuti afotokoze kuchuluka kwa maselo akufa ndi kuchuluka kwa ma cell.Countstar Rigel imapereka njira yachangu, yochulukira yowunikira magwiridwe antchito a GFP komanso kutheka nthawi imodzi.(Chithunzi 4)
Chithunzi 4: Maselo amapezeka pogwiritsa ntchito Hoechst 33342 (buluu) ndipo kuchuluka kwa ma cell a GFP (obiriwira) amatha kuzindikirika mosavuta.Maselo osatheka amakhala odetsedwa ndi propidium iodide (PI; red).
Viability ndi Kuwerengera Maselo
Kuwerengera kwa AO/PI Dual-fluoresces ndi mtundu woyeserera womwe umagwiritsidwa ntchito pozindikira kuchuluka kwa maselo, kutheka.Idagawika m'mawerengero a ma cell ndikuwerengera ma cell oyambira motengera mitundu yosiyanasiyana ya ma cell.The yankho lili osakaniza green-fluorescent nucleic acid banga, acridine lalanje, ndi redfluorescent nucleic acid banga, propidium iodide.Propidium iodide ndi utoto wopatula wa nembanemba womwe umangolowa m'maselo okhala ndi nembanemba wosokonekera pomwe acridine orange imalowa m'maselo onse mwa anthu.Mitundu yonse iwiri ikakhala mu phata, propidium iodide imayambitsa kuchepa kwa acridine orange fluorescence ndi fluorescence resonance energy transfer (FRET).Zotsatira zake, ma cell a nucleated okhala ndi nembanemba osawoneka bwino amadetsa zobiriwira za fulorosenti ndipo amawerengedwa kuti ndi amoyo, pomwe ma cell a nucleated okhala ndi nembanemba osokonekera amangopaka utoto wofiira wa fulorosenti ndipo amawerengedwa ngati akufa akamagwiritsa ntchito Countstar Rigel system.Zinthu zopanda ma nucleated monga maselo ofiira a magazi, mapulateleti ndi zinyalala sizimatuluka ndipo zimanyalanyazidwa ndi pulogalamu ya Countstar Rigel.(Chithunzi 5)
Chithunzi 5: Countstar yakonza njira yothimbirira yapawiri-fluorescence kuti ipezeke mosavuta, kutsimikiza kolondola kwa ndende ya PBMC ndi kutheka.Zitsanzo zodetsedwa ndi AO/PI zitha kuwunikidwa ndi Counstar Rigel
