(I-Flow cytometry , FCM ) isihlaziyi seseli esikala ukushuba kwe-fluorescence komaka amaseli anebala. Kuwubuchwepheshe obusezingeni eliphezulu obuthuthukisiwe obusekelwe ekuhlaziyweni nasekuhlungeni amaseli angawodwana. Ingakwazi ukulinganisa ngokushesha futhi ihlukanise usayizi, ukwakheka kwangaphakathi, i-DNA, i-RNA, amaprotheni, ama-antigen nezinye izakhiwo ezingokoqobo noma zamakhemikhali zamaseli, futhi ingasekelwe ekuqoqweni kwalezi zigaba.
I-cytometer egeleza ngokuyinhloko iqukethe izingxenye ezinhlanu ezilandelayo:
1 Igumbi lokugeleza kanye nesistimu ye-fluidics
2 Umthombo wokukhanya we-Laser kanye nesistimu yokubunjwa kwe-beam
3 Uhlelo lwe-Optical
4 I-elekthronikhi, isitoreji, isibonisi kanye nohlelo lokuhlaziya
5 Uhlelo lokuhlela amaseli
Phakathi kwazo, i-laser excitation emthonjeni wokukhanya we-laser kanye nesistimu yokwenza i-beam iyisilinganiso esiyinhloko samasiginali we-fluorescence ku-cytometry yokugeleza. Amandla okukhanya okuvusa amadlingozi kanye nesikhathi sokuchayeka kuhlobene nokuqina kwesignali ye-fluorescence. I-Laser iwumthombo wokukhanya ohambisanayo onganikeza ukukhanya kwegagasi elilodwa, ukuqina okuphezulu, nokukhanya okuzinze okuphezulu. Iwumthombo wokukhanya ojabulisayo ukuze uhlangabezane nalezi zidingo.
Kunamalensi amabili angama-cylindrical phakathi komthombo we-laser kanye negumbi lokugeleza. Lawa ma-lens agxilisa i-laser beam ene-cross-section eyindilinga ekhishwa emthonjeni we-laser ibe i-elliptical beam ene-cross-section encane (22 μm × 66 μm). Amandla e-laser angaphakathi kwalo mshayo oyi-elliptical asatshalaliswa ngokuya ngokusatshalaliswa okuvamile, okuqinisekisa ukuqina okungaguquki kokukhanya kwamaseli adlula endaweni yokuthola i-laser. Ngakolunye uhlangothi, isistimu ye-optical iqukethe amasethi amaningi amalensi, amaphinifa, nezihlungi, ezingahlukaniswa cishe ngamaqembu amabili: phezulu nomfula phansi kwegumbi lokugeleza.
Isistimu yokubona phambi kwegumbi lokugeleza iqukethe ilensi kanye nephinikhodi. Umsebenzi oyinhloko welensi nephinikhodi (imvamisa amalensi amabili kanye nephinifa) uwukugxilisa i-laser beam enesiphambano esiyindilinga esikhishwa umthombo we-laser sibe yi-elliptical nesigaba esiphambanayo esincane. Lokhu kusabalalisa amandla e-laser ngokuya ngokusatshalaliswa okuvamile, kuqinisekisa ukuqina kokukhanya okungaguquki kwamaseli kuyo yonke indawo yokuthola i-laser nokunciphisa ukuphazamiseka okuvela ekulahlekeni kokukhanya.
Kunezinhlobo ezintathu eziyinhloko zezihlungi:
1: Isihlungi se-Long pass (LPF) - sivumela kuphela ukukhanya okunamaza wamaza aphezulu kunevelu ethile ukuze kudlule kuwo.
2: Isihlungi se-Short-pass (SPF) - sivumela kuphela ukukhanya okunamaza wamaza angaphansi kwevelu ethile ukuthi kudlule.
3: Isihlungi se-bandpass (BPF) - sivumela kuphela ukukhanya ebangeni elithile le-wavelength ukuthi kudlule.
Inhlanganisela ehlukene yezihlungi ingaqondisa amasiginali e-fluorescence kumaza wamaza ahlukene kumashubhu e-photomultiplier (PMTs). Isibonelo, izihlungi zokuthola i-green fluorescence (FITC) phambi kwe-PMT yi-LPF550 ne-BPF525. Izihlungi ezisetshenziselwa ukuthola i-orange-red fluorescence (PE) phambi kwe-PMT yi-LPF600 ne-BPF575. Izihlungi zokuthola i-fluorescence ebomvu (CY5) phambi kwe-PMT yi-LPF650 ne-BPF675.
I-Flow cytometry isetshenziswa kakhulu ekuhlungeni amaseli. Ngokuthuthuka kobuchwepheshe bamakhompiyutha, ukuthuthukiswa kwe-immunology kanye nokusungulwa kobuchwepheshe be-monoclonal antibody, ukusetshenziswa kwayo ku-biology, imithi, ikhemisi kanye neminye imikhakha kuya ngokuya kwanda. Lezi zinhlelo zokusebenza zifaka phakathi ukuhlaziywa kokuguquguquka kweseli, i-cell apoptosis, ukuthayipha kwamaseli, ukuhlonza isimila, ukuhlaziya ukusebenza kwezidakamizwa, njll.
Isikhathi sokuthumela: Sep-21-2023
