Kev txheeb xyuas cov leukocytes hauv cov ntshav tag nrho yog ib qho kev kuaj mob niaj hnub hauv chaw kuaj mob lossis chaw txhab nyiaj ntshav.Qhov concentration thiab kev muaj peev xwm ntawm cov leukocytes yog qhov tseem ceeb index raws li kev tswj cov ntshav cia.Sib nrug los ntawm leukocyte, tag nrho cov ntshav muaj ntau cov platelets, cov qe ntshav liab, lossis cov khib nyiab ntawm tes, uas ua rau nws tsis tuaj yeem txheeb xyuas tag nrho cov ntshav ncaj qha rau hauv lub tshuab tsom lossis lub ntsej muag ci ntsa iab.Cov txheej txheem kev suav cov qe ntshav dawb koom nrog RBC lysis txheej txheem, uas yog siv sijhawm ntev.
AOPI Dual-fluoresces suav yog hom tshuaj ntsuam xyuas siv los kuaj xyuas cov cell concentration thiab muaj sia nyob.Cov tshuaj yog ua ke ntawm acridine txiv kab ntxwv (cov ntsuab-fluorescent nucleic acid stain) thiab propidium iodide (liab-fluorescent nucleic acid stain).Propidium iodide (PI) yog ib daim nyias nyias nyias dye uas tsuas yog nkag mus rau hauv cov hlwb nrog kev cuam tshuam daim nyias nyias, thaum acridine txiv kab ntxwv muaj peev xwm nkag mus rau tag nrho cov hlwb hauv cov pejxeem.Thaum ob lub dyes muaj nyob rau hauv lub nucleus, propidium iodide ua rau txo qis hauv acridine txiv kab ntxwv fluorescence los ntawm fluorescence resonance zog hloov (FRET).Raws li qhov tshwm sim, cov cell nucleated nrog cov kab mob tsis zoo stain fluorescent ntsuab thiab suav tias yog nyob, thaum cov cell nucleated nrog cov kab mob cuam tshuam tsuas yog stain fluorescent liab thiab suav tias tuag thaum siv Countstar® Rigel system.
Countstar Rigel yog ib qho kev daws teeb meem zoo tshaj plaws rau ntau qhov kev soj ntsuam ntawm cov pej xeem ntawm tes, pab kom ceev nrooj txheeb xyuas cov ntshav dawb hauv cov ntshav.
Cov txheej txheem sim:
1. Noj 20 µl ntawm cov ntshav kuaj thiab dilute cov qauv hauv 180 µl ntawm PBS.
2.Ntxiv 12µl AO/PI tov rau hauv 12µl qauv, maj mam tov nrog pipette;
3.Draw 20µl sib tov rau hauv chamber swb;
4.Cia cov hlwb los nyob hauv lub chamber li ntawm 1 feeb;
5.Insect tus swb rau hauv Countstar FL ntsuas;
6.Xaiv qhov kev ntsuam xyuas "AO/PI Viability", tom qab ntawd Sau Tus Qauv ID rau tus qauv no.
7.Xaiv Dilution piv, Cell Hom, nias 'Khiav' los pib qhov kev xeem.
Ceeb Toom: AO thiab PI yog cov muaj peev xwm carcinogen.Nws raug nquahu kom tus neeg teb xov tooj hnav cov cuab yeej tiv thaiv tus kheej (PPE) kom tsis txhob cuam tshuam ncaj qha rau ntawm daim tawv nqaij thiab qhov muag.
Qhov tshwm sim:
1. Bright Field Duab ntawm tag nrho cov ntshav
Nyob rau hauv daim duab kaj lug ntawm tag nrho cov ntshav, WBC's tsis pom ntawm cov ntshav liab.(Daim duab 1)
Daim duab 1 Bright daim duab ntawm tag nrho cov ntshav.
2. Fluorescence duab ntawm tag nrho cov ntshav
Cov AO thiab PI dye yog ob qho tib si stains DNA hauv cov cell nucleus ntawm cov hlwb.Yog li, Platelets, cov qe ntshav liab, lossis cov khib nyiab ntawm tes tsis tuaj yeem cuam tshuam rau cov leukocytes concentration thiab qhov tshwm sim muaj txiaj ntsig.Nyob leukocytes (ntsuab) thiab tuag leukocytes (Liab) tau yooj yim pom hauv cov duab fluorescence.(Daim duab 2)
Daim duab 2 Fluorescence duab ntawm tag nrho cov ntshav.(A).Duab ntawm AO Channel;(B) Duab ntawm PI Channel;(C) Ua ke cov duab ntawm AO thiab PI Channel.
3. Concentration thiab viability ntawm leukocytes
Lub Countstar FL software cia li suav cov cell ntawm peb chav chamber thiab suav cov nqi nruab nrab ntawm tag nrho WBC cell suav (1202), concentration (1.83 x 106 hlwb / ml), thiab % viability (82.04%).Tag nrho cov duab ntshav thiab cov ntaub ntawv tuaj yeem xa tawm tau yooj yim li PDF, Duab lossis Excel rau kev tshuaj xyuas ntxiv lossis khaws cov ntaub ntawv.
Daim duab 3 Screenshot ntawm Countstar Rigel Software
