Ukubala kwe-AOPI Dual-fluoresces wuhlobo lokuhlola olusetshenziselwa ukuthola ukugcwala kwamaseli nokusebenza.Isixazululo inhlanganisela ye-acridine orange (ibala eliluhlaza-fluorescent nucleic acid) kanye ne-propidium iodide (ibala elibomvu le-fluorescent nucleic acid).I-Propidium iodide (PI) wudayi wokukhishwa kwe-membrane ongena kuphela kumaseli anolwelwesi olusengozini, kuyilapho i-acridine orange ikwazi ukungena kuwo wonke amangqamuzana kubantu.Uma bobabili odayi bekhona ku-nucleus, i-propidium iodide ibangela ukuncipha kwe-acridine orange fluorescence ngokudluliswa kwe-fluorescence resonance energy (FRET).Ngenxa yalokho, amaseli angama-nucleated anolwelwesi olungaguquki angcolisa okuluhlaza okotshani futhi abalwa njengaphilayo, kuyilapho amaseli e-nucleated anolwelwesi olusengozini angcolisa kuphela okubomvu kwe-fluorescent futhi abalwa njengafile lapho kusetshenziswa uhlelo lwe-Countstar® FL.Izinto ezingezona i-nucleated njengamaseli egazi abomvu, ama-platelet kanye nemfucumfucu akushintshi futhi azinakwa isofthiwe ye-Countstar® FL.
Inqubo yokwelashwa kwe-Stem Cell
Umfanekiso 4 Ukuqapha ukusebenza nokubalwa kwamaseli we-mesenchymal stem cell (MSCs) ukuze asetshenziswe ekwelashweni kwamaseli.
Nquma ukusebenza kwe-MSC nge-AO/PI kanye ne-Trypan Blue assay
Umfanekiso 2. A. Isithombe se-MSC esingcoliswe yi-AO/PI kanye ne-Trypan Blue;2. Ukuqhathaniswa kwe-AO/PI nomphumela we-Trypan eluhlaza okwesibhakabhaka ngaphambi nangemuva kokuthutha.
Inkomba ye-cell refractive index iyashintsha, i-Trypan Blue staining yayingekho sobala kangako, kunzima ukunquma ukusebenza ngemuva kokuthutha.Nakuba i-fluorescence enemibala emibili ivumela ukungcoliswa kwamangqamuzana enucleated aphilayo nafile, okukhiqiza umphumela onembile wokuphila ngisho nalapho kukhona udoti, ama-platelet, namangqamuzana egazi abomvu.
