Qhov sib txawv tseem ceeb ntawm kev ncaj qha thiab tsis ncaj immunofluorescence yog qhov ncaj qha immunofluorescence siv ib qho tshuaj tiv thaiv kab mob uas ua haujlwm tawm tsam lub hom phiaj ntawm kev txaus siab thaum cov tshuaj tiv thaiv tsis ncaj ncees siv ob lub tshuaj tiv thaiv kab mob los sau lub hom phiaj ntawm kev txaus siab.
Immunofluorescence lossis cell imaging yog cov txheej txheem uas yog siv los sau cov hom phiaj antigen nrog fluorophore. Ntawm no, fluorophore yog ib qho tshuaj fluorescent uas tuaj yeem rov ua lub teeb thaum lub teeb pom kev zoo. Thaum lub fluorophore khi nrog lub hom phiaj antigen, nws tso cai rau kev tshawb pom ntawm lub hom phiaj molecule hauv cov qauv. Txhawm rau piav qhia ntxiv, thaum ib qho antigen khi nrog cov tshuaj tiv thaiv tshwj xeeb, nws tuaj yeem txuas nrog fluorophores. Yog li ntawd, nws yog ib qho yooj yim los soj ntsuam lub xub ntiag ntawm lub hom phiaj antigen nyob rau hauv cov qauv thaum soj ntsuam nyob rau hauv lub fluorescence microscope.
Dhau li ntawd, muaj ob hom immunofluorescence; direct thiab indirect immunofluorescence. Qhov txawv ntawm qhov ncaj thiab tsis ncaj immunofluorescence feem ntau yog nyob rau hauv tus naj npawb ntawm cov tshuaj tiv thaiv siv thiab cov fluorophore conjugation. Ntawd yog, hauv kev tiv thaiv kab mob ncaj qha, fluorophore ncaj qha txuas mus rau thawj cov tshuaj tiv thaiv kab mob thaum nyob rau hauv indirect immunofluorescence, fluorophore conjugate nrog cov tshuaj thib ob.
Direct Immunofluorescence yog dab tsi?
Immunofluorescence siv cov tshuaj tiv thaiv kab mob txhawm rau txheeb xyuas cov hom phiaj antigens. Direct immunofluorescence yog ib qho ntawm ob hom immunofluorescence. Hauv immunofluorescence ncaj qha, ib qho tshuaj tiv thaiv ib leeg (thawj cov tshuaj tiv thaiv kab mob) koom nrog thiab cov fluorophore ncaj qha txuas nrog thawj cov tshuaj tiv thaiv. Thaum khi cov tshuaj tiv thaiv kab mob nrog lub hom phiaj antigen, fluorophore emits fluorescence uas tuaj yeem kuaj pom los ntawm fluorescence microscope.
Daim duab 01: Direct Immunofluorescence
Txawm li cas los xij, cov tshuaj tiv thaiv kab mob ncaj qha yog ib txoj hauv kev kim vim tias thawj cov tshuaj tiv thaiv kab mob raug nqi ntau piv rau cov tshuaj tiv thaiv thib ob. Txawm li cas los xij, nws tsis cuam tshuam nrog cov kauj ruam ntxiv li no, cov txheej txheem luv dua. Tsis tas li ntawd, cov kev sib txuas tsis tshwj xeeb raug txo qis hauv cov immunofluorescence ncaj qha. Yog li ntawd, hom cross-reactivity yog tsawg. Tab sis nyob rau hauv kev kuaj pom, qhov rhiab heev ntawm cov tshuaj immunofluorescence ncaj qha tsis muaj zog nyob rau hauv kev sib piv rau indirect immunofluorescence.
Immunofluorescence Indirect yog dab tsi?
Indirect immunofluorescence yog hom thib ob ntawm immunofluorescence uas muaj ob hom tshuaj tiv thaiv xws li cov tshuaj tiv thaiv thawj thiab theem nrab hauv kev sau npe ntawm lub hom phiaj antigen. Hauv txoj kev no, fluorophore conjugates nrog cov tshuaj tiv thaiv kab mob thib ob. Li no, cov txheej txheem no suav nrog ib kauj ruam ntxiv.
Daim duab 02: Immunofluorescence
Txawm li cas los xij, qhov rhiab heev nyob rau hauv txoj kev no vim ntau cov fluorophores tuaj yeem sib txuas rau cov tshuaj tiv thaiv kab mob thib ob thiab nws ua rau kev tshawb pom yooj yim dua. Tsis tas li ntawd, indirect immunofluorescence tsis tshua kim vim qhov tseeb tias kev sib txuas ntawm cov tshuaj tiv thaiv kab mob thib ob tsis tshua kim thiab yooj yim dua. Nyob rau hauv kev sib piv rau qhov ncaj immunofluorescence, hom cross-reactivity yog siab nyob rau hauv txoj kev indirect.
Dab tsi zoo sib xws ntawm Direct thiab Indirect Immunofluorescence?
- Lawv yog ob hom txheej txheem immunofluorescence.
- Primary antibodies thiab fluorophore koom nrog ob txoj hauv kev.
- Tsis tas li, cov tshuaj tiv thaiv antigen-antibody tshwm sim hauv ob txoj hauv kev.
- Tsis tas li ntawd, ob qho tib si ncaj qha thiab tsis ncaj qha immunofluorescence siv lub fluorescence microscope rau kev kuaj pom cov tshuaj tiv thaiv.
Qhov txawv ntawm Direct thiab Indirect Immunofluorescence yog dab tsi?
Immunofluorescence tuaj yeem ncaj qha lossis tsis ncaj raws li fluorophore conjugation nrog cov tshuaj tiv thaiv siv. Hauv immunofluorescence ncaj qha, fluorophore conjugates nrog thawj cov tshuaj tiv thaiv, uas yog tib cov tshuaj tiv thaiv uas koom nrog hauv cov txheej txheem no. Nyob rau hauv sib piv rau qhov ntawd, indirect immunofluorescence, cov fluorophore conjugates nrog cov tshuaj tiv thaiv theem nrab, uas yog ib qho ntawm ob hom tshuaj tiv thaiv uas koom nrog hauv cov txheej txheem. Li no, qhov sib txawv tseem ceeb ntawm qhov ncaj thiab tsis ncaj immunofluorescence yog hom tshuaj tiv thaiv kab mob uas txuas nrog fluorophore.
Cov infographic hauv qab no muab ib sab ntawm kev sib piv ntawm qhov sib txawv ntawm qhov ncaj thiab tsis ncaj immunofluorescence.
Summary – Direct vs Indirect Immunofluorescence
Immunofluorescence yog cov txheej txheem uas siv los kuaj pom muaj cov tshuaj tiv thaiv tshwj xeeb hauv cov qauv. Cov txheej txheem no siv cov tshuaj tiv thaiv tshwj xeeb. Yog li ntawd, nyob rau hauv no, cov tshuaj tiv thaiv conjugate nrog fluorophores los xyuas lawv siv lub fluorescent microscope. Ntxiv mus, muaj ob hom immunofluorescence uas yog, ncaj qha thiab tsis ncaj immunofluorescence. Direct immunofluorescence muaj ib qho tshuaj tiv thaiv ib leeg thiab fluorophore ncaj qha sib txuas rau cov tshuaj tiv thaiv no. Indirect immunofluorescence muaj ob lub tshuaj tiv thaiv kab mob; thawj thiab theem nrab thiab fluorophore conjugated mus rau theem nrab antibody. Txij li ntau cov tshuaj tiv thaiv kab mob thib ob tuaj yeem khi nrog cov tshuaj tiv thaiv thawj zaug thiab ntau lub fluorophores tuaj yeem sib txuas nrog cov tshuaj tiv thaiv kab mob thib ob, kev tiv thaiv kab mob tsis ncaj yog ib txoj kev rhiab ntau dua li txoj kev ncaj nraim. Tsis tas li ntawd, txoj kev tsis ncaj ncees tsis tshua kim dua li txoj kev ncaj nraim. Yog li, qhov no yog qhov sib txawv ntawm qhov ncaj thiab tsis ncaj immunofluorescence.
