Tsarin Countstar yana haɗa sitometer hoto da counter ɗin tantanin halitta zuwa kayan aikin benci guda ɗaya.Wannan aikace-aikacen-kore, ƙarami, da tsarin siginar salula mai sarrafa kansa yana ba da mafita gabaɗaya don binciken ƙwayar cutar kansa, gami da ƙididdigewar kwayar halitta, yuwuwar (AO/PI, blue trypan), apoptosis (Annexin V-FITC/PI), tantanin halitta. sake zagayowar (PI), da GFP/RFP canja wuri.
Abstract
Ciwon daji na daya daga cikin manyan abubuwan da ke haddasa mutuwa a duniya, kuma samar da sabbin hanyoyin magance cutar kansa na da matukar muhimmanci.Kwayoyin ciwon daji shine ainihin abin bincike na ciwon daji, bayanai daban-daban suna buƙatar kimantawa daga kwayar cutar kansa.Wannan yanki na bincike yana buƙatar saurin bincike, abin dogaro, mai sauƙi da cikakken binciken tantanin halitta.Tsarin Countstar yana ba da dandamali mai sauƙi don nazarin ƙwayar cutar kansa.
Nazarin Ciwon Kankara Apoptosis na Countstar Rigel
Ana amfani da gwaje-gwajen Apoptosis akai-akai a cikin dakunan gwaje-gwaje da yawa don dalilai iri-iri daga tantance lafiyar al'adun tantanin halitta zuwa kimanta gubar rukunin mahadi.
Apoptosis assay nau'i ne da ake amfani dashi don tantance adadin apoptosis na sel ta hanyar Annexin V-FITC/PI.Annexin V yana ɗaure zuwa phosphatidylserine (PS) tare da farkon apoptosis cell ko necrosis cell.PI kawai yana shiga necrotic / sel apoptotic na zamani.(Hoto na 1)
A: Farkon apoptosis Annexin V (+), PI (-)
B: Late apoptosis Annexin V (+), PI (+)
Hoto1: Ƙara cikakkun bayanai na Hotunan Countstar Rigel (5 x haɓakawa) na sel 293, waɗanda aka bi da su tare da Annexin V FITC da PI
Nazarin Zagayowar Tantanin Halitta na Ciwon daji
Zagayowar tantanin halitta ko tsarin rarraba tantanin halitta shine jerin abubuwan da ke faruwa a cikin tantanin halitta wanda ke kaiwa ga rarrabawa da kwafin DNA ɗinsa (DNA replication) don samar da ƙwayoyin 'ya'ya mata biyu.A cikin sel tare da tsakiya, kamar yadda a cikin eukaryotes, tsarin tantanin halitta kuma ya kasu kashi uku: interphase, lokaci na mitotic (M), da cytokinesis.Propidium iodide (PI) wani rini ne na nukiliya wanda ake yawan amfani dashi don auna zagayowar tantanin halitta.Saboda rini ba zai iya shiga sel masu rai ba, ana gyara sel tare da ethanol kafin tabo.Duk sel ɗin suna tabo.Kwayoyin da ke shirin rarrabuwa za su ƙunshi adadin DNA da yawa kuma su nuna daidai gwargwadon ƙarfin haske.Ana amfani da bambance-bambance a cikin ƙarfin kyalli don tantance adadin sel a kowane lokaci na zagayowar tantanin halitta.Countstar na iya ɗaukar hoton kuma za a nuna sakamako a cikin software na FCS.(Hoto na 2)
Hoto 2: MCF-7 (A) da 293T (B) an lalata su tare da Kit ɗin Gano Kewar Tantanin halitta tare da PI, Countstar Rigel ne ya ƙaddara sakamakon, kuma an bincika ta FCS bayyana.
Yiwuwa da Ƙaddamar Canjawar GFP a cikin Tantanin halitta
A lokacin aikin bioprocess, GFP galibi ana amfani da shi don haɗa furotin da ke sake haɗawa azaman mai nuna alama.Ƙayyade mai kyalli na GFP na iya nuna maƙasudin furotin.Countstar Rigel yana ba da ƙima mai sauri da sauƙi don gwada canjin GFP har ma da yuwuwa.An lalata ƙwayoyin sel tare da Propidium iodide (PI) da Hoechst 33342 don ayyana matattun adadin tantanin halitta da jimillar yawan tantanin halitta.Countstar Rigel yana ba da hanya mai sauri, ƙididdigewa don kimanta ingancin magana da GFP a lokaci guda.(Hoto na 4)
Hoto 4: Kwayoyin suna samuwa ta amfani da Hoechst 33342 (blue) kuma ana iya ƙayyade yawan adadin GFP masu bayyanawa (kore).Kwayoyin da ba za a iya amfani da su ba suna da tabo da propidium iodide (PI; ja).
Yiwuwa da Ƙididdiga ta Tantanin halitta
AO/PI Dual-fluoresces ƙidayar ita ce nau'in tantancewar da ake amfani da ita don gano ƙwayar tantanin halitta, mai yiwuwa.Ya rabu zuwa kirga layin salula da kirgawa ta farko bisa ga nau'in tantanin halitta daban-daban.Maganin yana ƙunshe da haɗe-haɗe na koren-fluorescent nucleic acid tabo, acridine orange, da jajayen nucleic acid tabon, propidium iodide.Propidium iodide wani launi ne na cire membrane wanda ke shiga sel kawai tare da membranes masu rikitarwa yayin da acridine orange ke shiga duk sel a cikin yawan jama'a.Lokacin da rinannun biyu ke kasancewa a cikin tsakiya, propidium iodide yana haifar da raguwar acridine orange fluorescence ta hanyar canja wurin makamashi mai haske (FRET).Sakamakon haka, ƙwayoyin sel waɗanda ke da matattun membranes suna lalata kore mai kyalli kuma ana kirga su a matsayin masu rai, yayin da ƙwayoyin sel waɗanda ke da alaƙa da membranes kawai suna lalata ja kuma ana ƙidaya su a matsayin matattu lokacin amfani da tsarin Countstar Rigel.Abubuwan da ba su da ƙwayar cuta kamar ƙwayoyin jajayen jini, platelets da tarkace ba sa fitowa fili kuma software ɗin Countstar Rigel ta yi watsi da su.(Hoto na 5)
Hoto 5: Countstar ya inganta hanyar tabo mai haske-dual-fluorescence don sauƙi, daidaitaccen ƙayyadaddun ƙaddamarwar PBMC da yuwuwa.Za'a iya tantance samfuran da aka lalata da AO/PI tare da Countstar Rigel
