( Flow cytometry , FCM ) sisihlalutyi seseli esilinganisa ubunzulu be-fluorescence yeziphawuli zeeseli ezinebala. Bubuchwephesha obuphezulu obuphuhliswe ngokusekelwe kuhlalutyo kunye nokuhlelwa kweeseli enye. Iyakwazi ukulinganisa ngokukhawuleza kwaye ihlele ubungakanani, isakhiwo sangaphakathi, i-DNA, i-RNA, iiprotheni, i-antigens kunye nezinye izinto eziphathekayo okanye iikhemikhali zeeseli, kwaye zinokusekelwe ekuqokeleleni kwezi zigaba.
I-cytometer ehambayo iquka la macandelo amahlanu alandelayo:
1 Igumbi lokuhamba kunye nenkqubo ye-fluidics
2 Umthombo wokukhanya weLaser kunye nenkqubo yokubunjwa komqadi
3 Inkqubo yokubona
4 I-elektroniki, ukugcinwa, umboniso kunye nenkqubo yokuhlalutya
5 Inkqubo yokuhlela iiseli
Phakathi kwabo, ukuvuswa kwe-laser kumthombo wokukhanya kwe-laser kunye nenkqubo yokwenza i-beam ngumlinganiselo oyintloko weempawu ze-fluorescence kwi-cytometry yokuhamba. Ukuqina kokukhanya kokuvuselela kunye nexesha lokuvezwa lihambelana nokuqina komqondiso we-fluorescence. I-Laser ngumthombo wokukhanya ohambelanayo onokubonelela nge-wavelength enye, ukuqina okuphezulu, kunye nokukhanya okuphezulu kozinzo. Ngowona mthombo wokukhanya wovuselelo olufanelekileyo ukuhlangabezana nezi mfuno.
Kukho iilensi ezimbini zecylindrical phakathi komthombo we-laser kunye negumbi lokuhamba. Ezi lens zigxininisa i-laser beam ene-circular cross-section ephuma kumthombo we-laser ibe yintsimbi ye-elliptical ene-cross-section encinci (22 μm × 66 μm). Amandla e-laser ngaphakathi kwesi sithako se-elliptical sisasazwa ngokusasazwa okuqhelekileyo, ukuqinisekisa ukunyanzeliswa kokukhanya okungaguqukiyo kwiiseli ezidlula kwindawo yokufumanisa i-laser. Ngakolunye uhlangothi, inkqubo ye-optical iqulethe iiseti ezininzi zeelensi, i-pinholes, kunye nezihlungi, ezinokuthi zihlulwe ngokufanelekileyo zibe ngamaqela amabini: phezulu nangaphantsi kwegumbi lokuhamba.
Inkqubo ye-optical phambi kwegumbi lokuhamba liqukethe i-lens kunye ne-pinhole. Umsebenzi oyintloko we-lens kunye ne-pinhole (ngokuqhelekileyo iilensi ezimbini kunye ne-pinhole) kukugxila kwi-laser beam kunye ne-circular cross-section ekhutshwe ngumthombo we-laser ibe yi-elliptical beam kunye ne-cross-section encinci. Oku kusasaza amandla e-laser ngokokusasazwa okuqhelekileyo, kuqinisekisa ukuqina kokukhanya okungaguqukiyo kwiiseli kwindawo yokukhangela i-laser kunye nokunciphisa ukuphazamiseka ekukhanyeni okulahlekileyo.
Kukho iintlobo ezintathu eziphambili zezihluzo:
I-1: Isihluzi sokudlula ixesha elide (LPF) - sivumela kuphela ukukhanya okunobude obuphezulu kunexabiso elithile ukuba lidlule.
I-2: Isihluzo esifutshane (i-SPF) - sivumela ukukhanya kuphela ngamaza angaphantsi kwexabiso elithile ukuba kudlule.
I-3: Isihluzo se-Bandpass (BPF) - sivumela ukukhanya kuphela kuluhlu oluthile lwamaza ukuba ludlule.
Imidibaniso eyahlukeneyo yezihluzi inokwalathisa imiqondiso ye-fluorescence kubude bamaza ahlukeneyo kwiityhubhu zefotomultiplier (PMTs). Umzekelo, izihluzi zokubona i-fluorescence eluhlaza (FITC) phambi kwe-PMT yi-LPF550 kunye ne-BPF525. Izihluzi ezisetyenziselwa ukubona i-orenji-red fluorescence (PE) phambi kwe-PMT yi-LPF600 kunye ne-BPF575. Izihluzi zokubona i-fluorescence ebomvu (CY5) phambi kwe-PMT yi-LPF650 kunye ne-BPF675.
I-Flow cytometry isetyenziswa ikakhulu kukuhlelwa kweeseli. Ngenkqubela phambili yetekhnoloji yekhompyuter, uphuhliso lwe-immunology kunye nokuveliswa kwetekhnoloji ye-antibody ye-monoclonal, ukusetyenziswa kwayo kwi-biology, amayeza, ikhemesti kunye nezinye iindawo ziya zisanda. Ezi zicelo zibandakanya uhlalutyo lwe-cell dynamics, i-cell apoptosis, ukuchwetheza kweeseli, ukuxilongwa kwe-tumor, uhlalutyo lwe-chiza efficacy, njl.
Ixesha lokuposa: Sep-21-2023
