(Flow cytometry, FCM) ndi makina osanthula ma cell omwe amayesa mphamvu ya fluorescence ya zolembera zama cell.Ndiukadaulo wapamwamba kwambiri wopangidwa potengera kusanthula ndi kusanja ma cell amodzi.Imatha kuyeza ndikuyika mwachangu kukula, kapangidwe ka mkati, DNA, RNA, mapuloteni, ma antigen ndi zinthu zina zakuthupi kapena zamankhwala zama cell, ndipo zitha kutengera kusonkhanitsa kwa maguluwa.
Flow cytometer makamaka imakhala ndi magawo asanu awa:
1 Chipinda choyenda ndi dongosolo la fluidics
2 Gwero la kuwala kwa laser ndi njira yopangira matabwa
3 Optical system
4 Zipangizo zamagetsi, zosungirako, zowonetsera ndi kusanthula
5 Makina osankhira ma cell
Pakati pawo, chisangalalo cha laser mu gwero la kuwala kwa laser ndi dongosolo lopanga matabwa ndiye muyeso waukulu wa ma sigino a fluorescence mu flow cytometry.Kuchuluka kwa kuwala kwachisangalalo ndi nthawi yowonetsera zimagwirizana ndi mphamvu ya chizindikiro cha fluorescence.Laser ndi gwero lowunikira lomwe limatha kupereka mawonekedwe amtundu umodzi, kulimba kwambiri, komanso kuwunikira kokhazikika.Ndilo gwero loyatsira losangalatsa kuti likwaniritse izi.
Pali ma lens awiri a cylindrical pakati pa gwero la laser ndi chipinda choyenda.Magalasi awa amayang'ana mtengo wa laser wokhala ndi gawo lozungulira lochokera ku gwero la laser kupita kumtengo wozungulira wokhala ndi gawo laling'ono (22 μm × 66 μm).Mphamvu ya laser mkati mwa mtengo wa elliptical imagawidwa molingana ndi kugawidwa kwabwinobwino, kuwonetsetsa kuwunika kosasinthasintha kwa ma cell omwe amadutsa malo ozindikira laser.Kumbali inayi, mawonekedwe a kuwala amakhala ndi ma lens angapo, ma pinholes, ndi zosefera, zomwe zitha kugawidwa m'magulu awiri: kumtunda ndi kumunsi kwa chipinda choyenda.
Dongosolo loyang'ana kutsogolo kwa chipinda chothamanga chimakhala ndi lens ndi pinhole.Ntchito yayikulu ya mandala ndi pinhole (nthawi zambiri magalasi awiri ndi pinhole) ndikuwunikira mtengo wa laser wokhala ndi gawo lozungulira lopangidwa ndi gwero la laser mumtengo wozungulira wokhala ndi gawo laling'ono.Izi zimagawira mphamvu ya laser molingana ndi kugawidwa kwabwinobwino, kuwonetsetsa kuwunikira kosasinthasintha kwa ma cell kudera lozindikira la laser ndikuchepetsa kusokonezedwa ndi kuwala kosokera.
Pali mitundu itatu yayikulu ya zosefera:
1: Zosefera zazitali (LPF) - zimangolola kuwala kokhala ndi mafunde apamwamba kuposa mtengo wina wake kuti udutse.
2: Fyuluta ya Short-pass (SPF) - imangolola kuwala kokhala ndi mafunde otsika pansi pamtengo wapadera kuti udutse.
3: Bandpass fyuluta (BPF) - imangolola kuwala mumtundu wina wa wavelength kudutsa.
Zosefera zosiyanasiyana zimatha kuwongolera ma siginecha a fluorescence pamafunde osiyanasiyana kumachubu amtundu wa photomultiplier (PMTs).Mwachitsanzo, zosefera zozindikirira green fluorescence (FITC) kutsogolo kwa PMT ndi LPF550 ndi BPF525.Zosefera zomwe zimagwiritsidwa ntchito pozindikira orange-red fluorescence (PE) kutsogolo kwa PMT ndi LPF600 ndi BPF575.Zosefera zowonera red fluorescence (CY5) kutsogolo kwa PMT ndi LPF650 ndi BPF675.
Flow cytometry imagwiritsidwa ntchito makamaka posankha ma cell.Ndi kupita patsogolo kwaukadaulo wamakompyuta, chitukuko cha chitetezo chamthupi komanso kupangidwa kwaukadaulo wa monoclonal antibody, kugwiritsa ntchito kwake mu biology, mankhwala, pharmacy ndi magawo ena akuchulukirachulukira.Ntchitozi zikuphatikiza kusanthula kwama cell dynamics, cell apoptosis, kulemba ma cell, kuzindikira chotupa, kusanthula mphamvu ya mankhwala, ndi zina.
Nthawi yotumiza: Sep-21-2023
