I-Dual-fluorescence Viability(AO/PI), i-Acridine orange (AO) kanye ne-propidium iodide (PI) amabala enucleic nucleic kanye nodayi ababopha i-asidi.I-AO ingangena kulwelwesi lwamangqamuzana afile naphilayo futhi ingcolise i-nucleus, ikhiqize i-fluorescence eluhlaza.Ngokuphambene, i-PI ingangena kuphela kulwelwesi oluhlakazekayo lwamaseli ane-nucleated efile, ikhiqize i-fluorescence ebomvu.Ubuchwepheshe obusekelwe ezithombeni be-Countstar Rigel abubandakanyi izingcezu zamaseli, udoti, nezinhlayiya ze-artifact kanye nezehlakalo ezingaphansi kwama-platelet, okunikeza umphumela onembe kakhulu.Sengiphetha, uhlelo lwe-Countstar Rigel lungasetshenziswa kuso sonke isinyathelo senqubo yokukhiqiza amaseli.

I-T/NK Cell-Mediated Cytotoxicity, Ekwelapheni kwamaseli e-CAR-T okusanda kuvunywa yi-FDA, ama-T-lymphocyte akhiwe ngofuzo abopha ngokuqondile amangqamuzana omdlavuza (T) ahlosiwe futhi awabulale.Abahlaziyi be-Countstar Rigel bayakwazi ukuhlaziya le nqubo ephelele ye-T/NK Cell-Mediated Cytotoxicity.

Ucwaningo lwe-Cytotoxicity lwenziwa ngokubhala amaseli omdlavuza okuhlosiwe nge-CFSE noma ngokuwadlulisela nge-GFP.I-Hoechst 33342 ingase isetshenziselwe ukungcolisa wonke amaseli (womabili amaseli T namaseli wesimila).Kungenjalo, amaseli wesimila okuhlosiwe angangcoliswa nge-CFSE.I-Propidium iodide (PI) isetshenziselwa ukungcolisa amangqamuzana afile (womabili ama-T cell namaseli wesimila).Ukubandlulula phakathi kwamaseli ahlukene kungatholwa kusetshenziswa lelisu lokungcolisa amabala.

I-GFP Transfection Efficiency, Kuzakhi zofuzo zamangqamuzana, izinhlobo ezihlukahlukene zezinto eziphilayo, kanye ne-cell biology, isakhi sofuzo se-GFP sivame ukusetshenziswa njengentatheli ezifundweni zokukhuluma.Njengamanje, ososayensi bavamise ukusebenzisa izibonakhulu ezikhanyayo noma ama-fluorescent cytometers ukuze bahlaziye ukusebenza kahle kokudluliselwa kwamaseli ezincelisayo.Kodwa ukuphatha ubuchwepheshe obuyinkimbinkimbi be-cytometer yokugeleza okuthuthukisiwe kudinga u-opharetha onolwazi futhi oqeqeshwe kakhulu.I-Countstar Rigel inika amandla abasebenzisi ukuthi benze kalula futhi ngokunembile ukuhlola ukusebenza kahle kokudlulisela ngaphandle kwezindleko zokusebenza nezokulungisa ezihambisana ne-cytometry yokugeleza evamile.

I-Cell Apoptosis, Ukuqhubeka kwe-cell apoptosis kungagadwa kusetshenziswa i-FITC conjugated Annexin-V kuhlanganiswe ne-7-ADD.Izinsalela ze-Phosphatidylserine (PS) zivame ukutholakala engxenyeni engaphakathi ye-plasma membrane yamaseli anempilo.Ngesikhathi se-apoptosis yakuqala, ubuqotho be-membrane buyalahleka futhi i-PS izothuthelwa ngaphandle kolwelwesi lweseli.I-Annexin V inobudlelwane obuqinile ne-PS ngakho-ke iwumaka ofanelekile wamaseli okuqala e-apoptotic.

I-Cell Cycle, Ngesikhathi sokuhlukaniswa kwamaseli, amaseli aqukethe inani elikhulayo le-DNA.Ilebulwe nge-PI, ukwanda kokuqina kwe-fluorescence ngokulingana ngokuqondile nokuqoqwa kwe-DNA.Umehluko ekuqineni kwe-fluorescence yamangqamuzana awodwa yizinkomba zesimo sangempela somjikelezo weseli we-MCF 7 amaseli aphathwe nge-4μM ye-Nocodazole ukuze abophe lawa maseli ngezigaba ezahlukene zomjikelezo wabo weseli.Izithombe zenkundla ekhanyayo ezitholwe phakathi nalesi simo sokuhlola zisivumela ukuthi sihlonze iseli ngalinye.Ishaneli ye-PI fluorescence ye-Countstar Rigel ihlonza amasiginali e-DNA yeseli elilodwa ngisho nasemaqoqweni.Ukuhlaziywa okuningiliziwe kokuqina kwe-fluorescence kungenziwa kusetshenziswa i-FCS.

I-CD Marker Phenotyping, Izinhlobo ze-Countstar Rigel zinikeza indlela esheshayo, elula nezwela kakhulu, i-phenotyping esekelwe amasosha omzimba yamaseli esebenza kahle kakhulu.Ngezithombe ezinokulungiswa okuphezulu namandla okuhlaziya idatha ahlanganisiwe anamandla, i-Countstar Rigel ivumela abasebenzisi ukuthi bathole imiphumela ethembekile engashintshi ngaphandle kwesidingo sezilungiselelo zokulawula eziyinkimbinkimbi kanye nokulungiswa kwesinxephezelo se-fluorescence.

Ukuhlukaniswa kwamaseli we-Cytokine Induced Killer (CIK) kubonisa ikhwalithi yokusebenza evelele ye-Countstar Rigel analyzer ngokuqhathaniswa okuqondile nama-cytometer okugeleza okusezingeni eliphezulu.Ama-PBMC egundane esikweni agcotshwe nge-CD3-FITC, CD4-PE, CD8-PE, ne-CD56-PE, futhi enziwa yi-Interleukin (IL) 6. Bese ahlaziywa kanyekanye ne-Countstar® Rigel kanye ne-Flow Cytometry.Kulokhu kuhlolwa, i-CD3-CD4 , i-CD3-CD8, ne-CD3-CD56 zihlukaniswe ngamaqembu amathathu, ukuze kutholwe inani lamaseli ahlukene angaphansi.

Ukutholwa Kwamaseli Awohlokile nge-Immunofluorescence, amasosha omzimba e-Monoclonal akhiqiza imigqa yamaseli azolahlekelwa amanye ama-clones amahle phakathi nokwanda kwamaseli kanye nokudlula ngenxa yokuwohloka noma ukuguqulwa kofuzo.Ukulahlekelwa okuphezulu kuzothinta kakhulu ukukhiqizwa kwenqubo yokukhiqiza.Ukuqapha ukuwohloka kudlala indima ebalulekile ekulawuleni inqubo ukususa isivuno sama-antibody siye ezingeni eliphezulu.

Iningi lamasosha omzimba akhiqizwa embonini ye-BioPharma angatholwa ngokulebula kwe-immunofluorescence futhi ahlaziywe ngobuningi ngochungechunge lwe-Countstar Rigel.Izithombe zesiteshi se-bright-field ne-fluorescence ngezansi zibonisa ngokusobala lawo ma-clones alahlekelwe isibaluli sawo sokukhiqiza amasosha omzimba afiswayo.Ukuhlaziywa okunemininingwane eyengeziwe ngesofthiwe ye-DeNovo FCS Express Image kuyaqinisekisa, ukuthi u-86.35 % wawo wonke amaseli aveza ama-immunoglobulins, angu-3.34 % kuphela abonisa ukuthi akanayo.

I-Trypan (yenza ngofeleba B ngokuluhlaza okwesibhakabhaka) Ukubalwa kwamaseli, ukugcotshwa kwe-Trypan okuluhlaza okwesibhakabhaka kusasetshenziswa kumalebhu amaningi amasiko wamaseli.

I-Trypan Blue Viability kanye ne-Cell Density BioApp ingafakwa kuwo wonke amamodeli e-Countstar Rigel.Ama-algorithms ethu okubona isithombe avikelwe ahlaziya amapharamitha angaphezu kuka-20 ukuze ahlukanise into ngayinye etholiwe.

I-Cell Line Storage QC, Ekugcinweni kweseli, umqondo oyinkimbinkimbi wokuphatha ikhwalithi uqinisekisa ukuqapha okuphephile, okuphumelelayo kwayo yonke imikhiqizo yamaselula.Lokhu kuqinisekisa ikhwalithi ezinzile ye-cell cryopreserved, i-cryo-ilondolozelwe ukuhlolwa, ukuthuthukiswa kwenqubo, nokukhiqizwa.

I-Countstar Rigel ithola izithombe ezinokulungiswa okuphezulu, ihlaziya izici ezihlukahlukene ze-morphological zezinto zeselula ezifana nobubanzi, ukwakheka, nokuthambekela kokuhlanganisa.Izithombe zezinyathelo ezihlukene zenqubo zingaqhathaniswa kalula komunye nomunye.Ngakho-ke ukuhlukahluka komumo nokuhlanganisa kungatholwa kalula, ngokugwema izilinganiso zomuntu.Futhi isizindalwazi se-Countstar Rigel sinohlelo lokuphatha oluyinkimbinkimbi lokugcina nokubuyiswa kwezithombe nedatha.