(Flow cytometry, FCM) nisesengura rya selile ipima ubukana bwa fluorescence yibimenyetso bya selile.Nubuhanga buhanga buhanitse bwateye imbere bushingiye kubisesengura no gutondekanya selile imwe.Irashobora gupima byihuse no gutondekanya ingano, imiterere y'imbere, ADN, RNA, proteyine, antigene nibindi bintu bifatika cyangwa imiti ya selile, kandi birashobora gushingira ku ikusanyirizo ryibi byiciro.
Flow cytometer igizwe ahanini nibice bitanu bikurikira:
1 Flow chamber na sisitemu ya fluidics
2 Inkomoko yumucyo na sisitemu yo gushiraho ibiti
Sisitemu nziza
4 Ibyuma bya elegitoroniki, kubika, kwerekana no gusesengura sisitemu
Sisitemu yo gutondekanya selile
Muri byo, kwishimisha lazeri mumasoko yumucyo wa laser hamwe na sisitemu yo gushiraho urumuri nicyo gipimo nyamukuru cyibimenyetso bya fluorescence muri cytometrike.Ubukomezi bwurumuri rwo kwishima nigihe cyo kumurika bifitanye isano nuburemere bwikimenyetso cya fluorescence.Laser nisoko yumucyo ishobora gutanga umurongo umwe-mwinshi, ubukana bwinshi, hamwe no kumurika cyane.Nibintu byiza bitanga urumuri rwo kwuzuza ibisabwa.
Hano hari lens ebyiri zibiri hagati yisoko ya laser nicyumba gitemba.Izi linzira zibanda kumurongo wa lazeri hamwe nuruziga ruzengurutse ruva mumasoko ya laser mumurongo wa elliptique hamwe nuduce duto duto (22 μm × 66 μ m).Ingufu za lazeri muri iki kibaho cya elliptique zitangwa hakurikijwe isaranganya risanzwe, zitanga imbaraga zihoraho zo kumurika ingirabuzimafatizo zinyura ahantu hagaragara.Kurundi ruhande, sisitemu ya optique igizwe nibice byinshi bya lens, pinholes, na filtri, bishobora kugabanywa mubice bibiri: hejuru no hepfo yicyumba gitemba.
Sisitemu ya optique imbere yicyumba gitemba igizwe na lens na pinhole.Igikorwa nyamukuru cya lens na pinhole (mubisanzwe lens ebyiri na pinhole) nukwibanda kumurongo wa lazeri hamwe nuruziga ruzengurutse rwasohowe nisoko ya laser mumurongo wa elliptique hamwe nuduce duto duto.Ibi bikwirakwiza ingufu za lazeri ukurikije isaranganya risanzwe, ikemeza ko urumuri rudasanzwe rwimikorere ya selile ahantu hagaragara kandi hagabanywa intambamyi zituruka kumucyo wayobye.
Hariho ubwoko butatu bwingenzi bwa filteri:
1: Inzira ndende ya filteri (LPF) - yemerera gusa urumuri rufite uburebure burenze agaciro runaka kunyuramo.
2: Bigufi-byungurura (SPF) - byemerera gusa urumuri nuburebure bwumurongo munsi yagaciro kanyuze.
3: Akayunguruzo ka bande (BPF) - yemerera gusa urumuri mumurongo wihariye wumurongo unyuramo.
Ihuriro ritandukanye ryayunguruzo rishobora kuyobora ibimenyetso bya fluorescence kumurambararo utandukanye kumiyoboro ya Photomultiplier (PMTs).Kurugero, muyunguruzi kugirango tumenye icyatsi kibisi (FITC) imbere ya PMT ni LPF550 na BPF525.Akayunguruzo gakoreshwa mu kumenya orange-umutuku fluorescence (PE) imbere ya PMT ni LPF600 na BPF575.Akayunguruzo ko kumenya fluorescence itukura (CY5) imbere ya PMT ni LPF650 na BPF675.
Flow cytometrie ikoreshwa cyane mugutondekanya selile.Iterambere ry’ikoranabuhanga rya mudasobwa, iterambere ry’ikingira no guhanga ikoranabuhanga rya antibody ya monoclonal, ikoreshwa ry’ibinyabuzima, ubuvuzi, farumasi n’izindi nzego riragenda ryiyongera.Izi porogaramu zirimo isesengura ryimikorere ya selile, selile apoptose, kwandika selile, gusuzuma ibibyimba, gusesengura imikorere yibiyobyabwenge, nibindi.
Igihe cyo kohereza: Nzeri-21-2023
